Inhibitors of bruton&#39;s tyrosine kinase

ABSTRACT

Disclosed herein are amido compounds that form covalent bonds with Bruton&#39;s tyrosine kinase (Btk). Also described are irreversible inhibitors of Btk. Also disclosed are pharmaceutical compositions that include the compounds. Methods of using the Btk inhibitors are disclosed, alone or in combination with other therapeutic agents, for the treatment of autoimmune diseases or conditions, heteroimmune diseases or conditions, cancer, including lymphoma, and inflammatory diseases or conditions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/032,257, filed Aug. 1, 2014, and U.S. Provisional Application No.62/199,108, filed Jul. 30, 2015, and both of said applications areincorporated herein by reference in their entireties.

FIELD OF THE INVENTION

Described herein are compounds, methods of making such compounds,pharmaceutical compositions and medicaments containing such compounds,and methods of using such compounds and compositions to inhibit theactivity of tyrosine kinases.

BACKGROUND OF THE INVENTION

Bruton's tyrosine kinase (Btk), a member of the Tec family ofnon-receptor tyrosine kinases, is a key signaling enzyme expressed inall hematopoietic cells types except T lymphocytes and natural killercells. Btk plays an essential role in the B-cell signaling pathwaylinking cell surface B-cell receptor (BCR) stimulation to downstreamintracellular responses.

Btk is a key regulator of B-cell development, activation, signaling, andsurvival (Kurosaki, Curr Op Imm, 2000, 276-281; Schaeffer andSchwartzberg, Curr Op Imm 2000, 282-288). In addition, Btk plays a rolein a number of other hematopoetic cell signaling pathways, e.g., Tolllike receptor (TLR) and cytokine receptor-mediated TNF-α production inmacrophages, IgE receptor (FcepsilonRI) signaling in Mast cells,inhibition of Fas/APO-1 apoptotic signaling in B-lineage lymphoid cells,and collagen-stimulated platelet aggregation. See, e.g., C. A. Jeffries,et al., (2003), Journal of Biological Chemistry 278:26258-26264; N. J.Horwood, et al., (2003), The Journal of Experimental Medicine197:1603-1611; Iwaki et al. (2005), Journal of Biological Chemistry280(48):40261-40270; Vassilev et al. (1999), Journal of BiologicalChemistry 274(3):1646-1656, and Quek et al. (1998), Current Biology8(20):1137-1140.

SUMMARY OF THE INVENTION

Described herein are inhibitors of Bruton's tyrosine kinase (Btk). Alsodescribed herein are irreversible inhibitors of Btk.

Further described are irreversible inhibitors of Btk that form acovalent bond with a cysteine residue on Btk. Further described hereinare irreversible inhibitors of other tyrosine kinases, wherein the othertyrosine kinases share homology with Btk by having a cysteine residue(including a Cys 481 residue) that can form a covalent bond with theirreversible inhibitor (such tyrosine kinases, are referred herein as“Btk tyrosine kinase cysteine homologs”).

Further described are irreversible inhibitors of Btk that form acovalent bond with a serine residue on C481S mutated Btk. Specificallydescribed are irreversible inhibitors of Btk that form a covalent bondwith a serine481 residue on C481S mutated Btk (Woyach, et al. Resistancemechanisms for the Bruton's tyrosine kinase inhibitor ibrutinib, N EnglJ Med. 2014, 12; 370(24):2286-94). Further described herein areirreversible inhibitors of other tyrosine kinases, wherein the othertyrosine kinases share homology with C481S mutated Btk by having aserine residue (including a homologous residue to BTK C481S residue)that can form a covalent bond with the inhibitor.

Also described herein are methods for synthesizing such irreversibleinhibitors, methods for using such irreversible inhibitors in thetreatment of diseases (including diseases wherein irreversibleinhibition of Btk provides therapeutic benefit to a patient having thedisease). Further described are pharmaceutical compositions that includean irreversible inhibitor of Btk.

Thus, in one specific aspect, the present invention provides methods forpreventing, treating or ameliorating in a mammal a disease or conditionthat is causally related to the aberrant activity of a tyrosine kinasereceptor in vivo, which comprises administering to the mammal aneffective disease-treating or condition-treating amount of a compoundaccording to Formula (I) having the structure:

wherein:

-   A is

-   Hy is 2-pyridyl substituted with 1-5 groups independently selected    from R⁴, or-   Hy is imidazolyl, thiazolyl, oxazolyl, pyrazolyl, oxadiazolyl,    thiadiazolyl, triazolyl, isothiazolyl, or tetrazolyl,-   each of which substituted with 1-2 groups independently selected    from R⁴;-   each R¹ and R² is independently H, alkyl, or CN; or R¹ and R²    together form a bond; R³ is independently H, alkyl, CN, or    cycloalkyl;-   each R⁴ is independently H, halo, hydroxyl, CN, substituted or    unsubstituted alkyl, substituted or unsubstituted amino, substituted    or unsubstituted alkoxy, substituted or unsubstituted amido,    substituted or unsubstituted sulfonyl, substituted or unsubstituted    carboxy, substituted or unsubstituted aryl, or substituted or    unsubstituted heteroaryl; or two R⁴s connect together with Hy to    form a bicyclic ring;-   each n is independently 0, 1, or 2; and-   p is 0, 1 or 2;-   or a metabolite, a solvate, a pharmaceutically acceptable salt, or a    prodrug thereof.

In some embodiments, Hy is 2-pyridyl substituted with 1-5 groupsindependently selected from R⁴, or Hy is imidazolyl, thiazolyl,oxazolyl, pyrazolyl, oxadiazolyl, thiadiazolyl, triazolyl, each of whichsubstituted with 1-2 groups independently selected from R⁴, each R⁴ isindependently H, halo, hydroxyl, CN, substituted or unsubstituted alkyl,substituted or unsubstituted amino, substituted or unsubstituted alkoxy,substituted or unsubstituted amido, substituted or unsubstitutedsulfonyl, substituted or unsubstituted carboxy, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl.

In one embodiment, the tyrosine kinase receptor is Btk receptor.

In one embodiment, the active site is a cavity in which the compound orthe moiety binds to the tyrosine kinase.

In one embodiment, the disease or condition is an autoimmune disease, aheteroimmune disease, a cancer, mastocytosis, osteoporosis or boneresorption disorder, or an inflammatory disease.

In another aspect, the present invention provides a compound accordingto Formula (I) having the structure:

wherein:

-   A is

-   Hy is 2-pyridyl substituted with 1-5 groups independently selected    from R⁴, or-   Hy is imidazolyl, thiazolyl, oxazolyl, pyrazolyl, oxadiazolyl,    thiadiazolyl, triazolyl, isothiazolyl, or tetrazolyl, each of which    substituted with 1-2 groups independently selected from R⁴;-   each R¹ and R² is independently H, alkyl, or CN; or R¹ and R²    together form a bond;-   R³ is independently H, alkyl, CN, or cycloalkyl;-   each R⁴ is independently H, halo, hydroxyl, CN, substituted or    unsubstituted alkyl, substituted or unsubstituted amino, substituted    or unsubstituted alkoxy, substituted or unsubstituted amido,    substituted or unsubstituted sulfonyl, substituted or unsubstituted    carboxy, substituted or unsubstituted aryl, or substituted or    unsubstituted heteroaryl; or two adjacent R⁴s connect together with    Hy to form a bicyclic ring optionally substituted with alkyl,    haloalkyl or CN;-   each n is independently 0, 1, or 2; and-   p is 0, 1 or 2;-   or a metabolite, a solvate, a pharmaceutically acceptable salt, or a    prodrug thereof.

In another aspect, the present invention provides a compound accordingto Formula (I) having the structure:

wherein:

-   A is

-   Hy is 2-pyridyl substituted with 1-5 groups independently selected    from R⁴, or-   Hy is imidazolyl, thiazolyl, oxazolyl, pyrazolyl, oxadiazolyl,    thiadiazolyl, triazolyl, each of which substituted with 1-2 groups    independently selected from R⁴;-   each R¹ and R² is independently H, alkyl, or CN; or R¹ and R²    together form a bond;-   R³ is independently H, alkyl, CN, or cycloalkyl;-   each R⁴ is independently H, halo, hydroxyl, CN, substituted or    unsubstituted alkyl, substituted or unsubstituted amino, substituted    or unsubstituted alkoxy, substituted or unsubstituted amido,    substituted or unsubstituted sulfonyl, substituted or unsubstituted    carboxy, substituted or unsubstituted aryl, or substituted or    unsubstituted heteroaryl;-   and each n is independently 0, 1, or 2;-   or a metabolite, a solvate, a pharmaceutically acceptable salt, or a    prodrug thereof.

In some embodiments, Hy is selected from:

In a particular embodiment, the compound is:

and wherein R³, R⁴, and n are as described herein.

In another aspect, the present invention provides pharmaceuticalcompositions comprising a therapeutically effective amount of a compoundof Formula (I) and a pharmaceutically acceptable excipient. In oneembodiment, the pharmaceutical composition comprising the compound ofFormula (I) is formulated for a route of administration selected fromoral administration, parenteral administration, buccal administration,nasal administration, topical administration, or rectal administration.In another aspect is a method for treating an autoimmune disease orcondition comprising administering to a patient in need atherapeutically effective amount of a compound of Formula (I). In oneembodiment the autoimmune disease is selected from rheumatoid arthritisor lupus. In a further aspect is a method for treating a heteroimmunedisease or condition comprising administering to a patient in need atherapeutically effective amount of a compound of Formula (I). In yetanother embodiment is a method for treating a cancer comprisingadministering to a patient in need a therapeutically effective amount ofa compound of Formula (I). In one embodiment the cancer is a B-cellproliferative disorder.

In another embodiment the B-cell proliferative disorder is diffuse largeB cell lymphoma, follicular lymphoma or chronic lymphocytic leukemia.

Yet a further aspect is a method for treating mastocytosis comprisingadministering to a patient in need a therapeutically effective amount ofa compound of Formula (I).

Another aspect is a method for treating osteoporosis or bone resorptiondisorders comprising administering to a patient in need atherapeutically effective amount of a compound of Formula (I).

A further aspect is a method for treating an inflammatory disease orcondition comprising administering to a patient in need atherapeutically effective amount of a compound of Formula (I).

Any combination of the groups described above for the various variablesis contemplated herein. It is understood that substituents andsubstitution patterns on the compounds provided herein can be selectedby one of ordinary skill in the art to provide compounds that arechemically stable and that can be synthesized by techniques known in theart, as well as those set forth herein.

In a further aspect are provided pharmaceutical compositions, whichinclude a therapeutically effective amount of at least one of any of thecompounds herein, or a pharmaceutically acceptable salt,pharmaceutically active metabolite, pharmaceutically acceptable prodrug,or pharmaceutically acceptable solvate. In certain embodiments,compositions provided herein further include a pharmaceuticallyacceptable diluent, excipient and/or binder.

Pharmaceutical compositions formulated for administration by anappropriate route and means containing effective concentrations of oneor more of the compounds provided herein, or pharmaceutically effectivederivatives thereof, that deliver amounts effective for the treatment,prevention, or amelioration of one or more symptoms of diseases,disorders or conditions that are modulated or otherwise affected bytyrosine kinase activity, or in which tyrosine kinase activity isimplicated, are provided. The effective amounts and concentrations areeffective for ameliorating any of the symptoms of any of the diseases,disorders or conditions disclosed herein.

In certain embodiments, provided herein is a pharmaceutical compositioncontaining: i) a physiologically acceptable carrier, diluent, and/orexcipient; and ii) one or more compounds provided herein.

In one aspect, provided herein are methods for treating a patient byadministering a compound provided herein. In some embodiments, providedherein is a method of inhibiting the activity of tyrosine kinase(s),such as Btk, or of treating a disease, disorder, or condition, whichwould benefit from inhibition of tyrosine kinase(s), such as Btk, in apatient, which includes administering to the patient a therapeuticallyeffective amount of at least one of any of the compounds herein, orpharmaceutically acceptable salt, pharmaceutically active metabolite,pharmaceutically acceptable prodrug, or pharmaceutically acceptablesolvate.

In another aspect, provided herein is the use of a compound disclosedherein for inhibiting Bruton's tyrosine kinase (Btk) activity or for thetreatment of a disease, disorder, or condition, which would benefit frominhibition of Bruton's tyrosine kinase (Btk) activity.

In some embodiments, compounds provided herein are administered to ahuman.

In some embodiments, compounds provided herein are orally administered.

In other embodiments, compounds provided herein are used for theformulation of a medicament for the inhibition of tyrosine kinaseactivity. In some other embodiments, compounds provided herein are usedfor the formulation of a medicament for the inhibition of Bruton'styrosine kinase (Btk) activity.

Articles of manufacture including packaging material, a compound orcomposition or pharmaceutically acceptable derivative thereof providedherein, which is effective for inhibiting the activity of tyrosinekinase(s), such as Btk, within the packaging material, and a label thatindicates that the compound or composition, or pharmaceuticallyacceptable salt, pharmaceutically active metabolite, pharmaceuticallyacceptable prodrug, or pharmaceutically acceptable solvate thereof, isused for inhibiting the activity of tyrosine kinase(s), such as Btk, areprovided.

In a further aspect, provided herein is a method for inhibiting Bruton'styrosine kinase in a subject in need thereof by administering to thesubject thereof a composition containing a therapeutically effectiveamount of at least one compound having the structure of Formula (I). Insome embodiments, the subject in need is suffering from an autoimmunedisease, e.g., inflammatory bowel disease, arthritis, lupus, rheumatoidarthritis, psoriatic arthritis, osteoarthritis, Still's disease,juvenile arthritis, diabetes, myasthenia gravis, Hashimoto'sthyroiditis, Ord's thyroiditis, Graves' disease Sjögren's syndrome,multiple sclerosis, Guillain-Barré syndrome, acute disseminatedencephalomyelitis, Addison's disease, opsoclonus-myoclonus syndrome,ankylosing spondylitis, antiphospholipid antibody syndrome, aplasticanemia, autoimmune hepatitis, coeliac disease, Goodpasture's syndrome,idiopathic thrombocytopenic purpura, optic neuritis, scleroderma,primary biliary cirrhosis, Reiter's syndrome, Takayasu's arteritis,temporal arteritis, warm autoimmune hemolytic anemia, Wegener'sgranulomatosis, psoriasis, alopecia universalis, Behçet's disease,chronic fatigue, dysautonomia, endometriosis, interstitial cystitis,neuromyotonia, scleroderma, or vulvodynia.

In other embodiments, the subject in need is suffering from aheteroimmune condition or disease, e.g., graft versus host disease,transplantation, transfusion, anaphylaxis, allergy, type Ihypersensitivity, allergic conjunctivitis, allergic rhinitis, or atopicdermatitis.

In certain embodiments, the subject in need is suffering from aninflammatory disease, e.g., asthma, appendicitis, blepharitis,bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis,cholecystitis, colitis, conjunctivitis, cystitis, dacryoadenitis,dermatitis, dermatomyositis, encephalitis, endocarditis, endometritis,enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis,fibrositis, gastritis, gastroenteritis, hepatitis, hidradenitissuppurativa, laryngitis, mastitis, meningitis, myelitis myocarditis,myositis, nephritis, oophoritis, orchitis, osteitis, otitis,pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis,pleuritis, phlebitis, pneumonitis, pneumonia, proctitis, prostatitis,pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis,tendonitis, tonsillitis, uveitis, vaginitis, vasculitis, or vulvitis.

In further embodiments, the subject in need is suffering from a cancer.In one embodiment, the cancer is a B-cell proliferative disorder, e.g.,diffuse large B cell lymphoma, follicular lymphoma, chronic lymphocyticlymphoma, chronic lymphocytic leukemia, B-cell prolymphocytic leukemia,lymphoplasmacytic lymphoma/Waldenström macroglobulinemia, splenicmarginal zone lymphoma, plasma cell myeloma, plasmacytoma, extranodalmarginal zone B cell lymphoma, nodal marginal zone B cell lymphoma,mantle cell lymphoma, mediastinal (thymic) large B cell lymphoma,intravascular large B cell lymphoma, primary effusion lymphoma, burkittlymphoma/leukemia, or lymphomatoid granulomatosis. In some embodiments,where the subject is suffering from a cancer, an anti-cancer agent isadministered to the subject in addition to one of the above-mentionedcompounds. In one embodiment, the anti-cancer agent is an inhibitor ofmitogen-activated protein kinase signaling, e.g., U0126, PD98059,PD184352, PD0325901, ARRY-142886, SB239063, SP600125, BAY 43-9006,wortmannin, or LY294002.

In further embodiments, the subject in need is suffering from athromboembolic disorder, e.g., myocardial infarct, angina pectoris,reocclusion after angioplasty, restenosis after angioplasty, reocclusionafter aortocoronary bypass, restenosis after aortocoronary bypass,stroke, transitory ischemia, a peripheral arterial occlusive disorder,pulmonary embolism, or deep venous thrombosis.

In a further aspect, provided herein is a method for treating anautoimmune disease by administering to a subject in need thereof acomposition containing a therapeutically effective amount of at leastone compound having the structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). In one embodiment, the autoimmune disease is arthritis.In another embodiment, the autoimmune disease is lupus. In someembodiments, the autoimmune disease is inflammatory bowel disease(including Crohn's disease and ulcerative colitis), rheumatoidarthritis, psoriatic arthritis, osteoarthritis, Still's disease,juvenile arthritis, lupus, diabetes, myasthenia gravis, Hashimoto'sthyroiditis, Ord's thyroiditis, Graves' disease Sjögren's syndrome,multiple sclerosis, Guillain-Barré syndrome, acute disseminatedencephalomyelitis, Addison's disease, opsoclonus-myoclonus syndrome,ankylosing spondylitis, antiphospholipid antibody syndrome, aplasticanemia, autoimmune hepatitis, coeliac disease, Goodpasture's syndrome,idiopathic thrombocytopenic purpura, optic neuritis, scleroderma,primary biliary cirrhosis, Reiter's syndrome, Takayasu's arteritis,temporal arteritis, warm autoimmune hemolytic anemia, Wegener'sgranulomatosis, psoriasis, alopecia universalis, Behçet's disease,chronic fatigue, dysautonomia, endometriosis, interstitial cystitis,neuromyotonia, scleroderma, or vulvodynia.

In a further aspect, provided herein is a method for treating aheteroimmune condition or disease by administering to a subject in needthereof a composition containing a therapeutically effective amount ofat least one compound having the structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). In some embodiments, the heteroimmune condition ordisease is graft versus host disease, transplantation, transfusion,anaphylaxis, allergy, type I hypersensitivity, allergic conjunctivitis,allergic rhinitis, or atopic dermatitis.

In a further aspect, provided herein is a method for treating aninflammatory disease by administering to a subject in need thereof acomposition containing a therapeutically effective amount of at leastone compound having the structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). In some embodiments, the inflammatory disease is asthma,inflammatory bowel disease (including Crohn's disease and ulcerativecolitis), appendicitis, blepharitis, bronchiolitis, bronchitis,bursitis, cervicitis, cholangitis, cholecystitis, colitis,conjunctivitis, cystitis, dacryoadenitis, dermatitis, dermatomyositis,encephalitis, endocarditis, endometritis, enteritis, enterocolitis,epicondylitis, epididymitis, fasciitis, fibrositis, gastritis,gastroenteritis, hepatitis, hidradenitis suppurativa, laryngitis,mastitis, meningitis, myelitis myocarditis, myositis, nephritis,oophoritis, orchitis, osteitis, otitis, pancreatitis, parotitis,pericarditis, peritonitis, pharyngitis, pleuritis, phlebitis,pneumonitis, pneumonia, proctitis, prostatitis, pyelonephritis,rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendonitis,tonsillitis, uveitis, vaginitis, vasculitis, or vulvitis.

In yet another aspect, provided herein is a method for treating a cancerby administering to a subject in need thereof a composition containing atherapeutically effective amount of at least one compound having thestructure of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh). In one embodiment,the cancer is a B-cell proliferative disorder, e.g., diffuse large Bcell lymphoma, follicular lymphoma, chronic lymphocytic lymphoma,chronic lymphocytic leukemia, B-cell prolymphocytic leukemia,lymphoplasmacytic lymphoma/Waldenström macroglobulinemia, splenicmarginal zone lymphoma, plasma cell myeloma, plasmacytoma, extranodalmarginal zone B cell lymphoma, nodal marginal zone B cell lymphoma,mantle cell lymphoma, mediastinal (thymic) large B cell lymphoma,intravascular large B cell lymphoma, primary effusion lymphoma, burkittlymphoma/leukemia, or lymphomatoid granulomatosis. In some embodiments,where the subject is suffering from a cancer, an anti-cancer agent isadministered to the subject in addition to one of the above-mentionedcompounds. In one embodiment, the anti-cancer agent is an inhibitor ofmitogen-activated protein kinase signaling, e.g., U0126, PD98059,PD184352, PD0325901, ARRY-142886, SB239063, SP600125, BAY 43-9006,wortmannin, or LY294002.

In another aspect, provided herein is a method for treating athromboembolic disorder by administering to a subject in need thereof acomposition containing a therapeutically effective amount of at leastone compound having the structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). In some embodiments, the thromboembolic disorder ismyocardial infarct, angina pectoris, reocclusion after angioplasty,restenosis after angioplasty, reocclusion after aortocoronary bypass,restenosis after aortocoronary bypass, stroke, transitory ischemia, aperipheral arterial occlusive disorder, pulmonary embolism, or deepvenous thrombosis.

In a further aspect, provided herein is a method for treating anautoimmune disease by administering to a subject in need thereof acomposition containing a therapeutically effective amount of a compoundthat forms a covalent bond with Bruton's tyrosine kinase. In oneembodiment, the compound forms a covalent bound with the activated formof Bruton's tyrosine kinase. In further or alternative embodiments, thecompound irreversibly inhibits the Bruton's tyrosine kinase to which itis covalently bound. In a further or alternative embodiment, thecompound forms a covalent bond with a cysteine residue on Bruton'styrosine kinase.

In a further aspect, provided herein is a method for treating aheteroimmune condition or disease by administering to a subject in needthereof a composition containing a therapeutically effective amount of acompound that forms a covalent bond with Bruton's tyrosine kinase. Inone embodiment, the compound forms a covalent bound with the activatedform of Bruton's tyrosine kinase. In further or alternative embodiments,the compound irreversibly inhibits the Bruton's tyrosine kinase to whichit is covalently bound. In a further or alternative embodiment, thecompound forms a covalent bond with a cysteine residue on Bruton'styrosine kinase.

In a further aspect, provided herein is a method for treating aninflammatory disease by administering to a subject in need thereof acomposition containing a therapeutically effective amount of a compoundthat forms a covalent bond with Bruton's tyrosine kinase. In oneembodiment, the compound forms a covalent bound with the activated formof Bruton's tyrosine kinase. In further or alternative embodiments, thecompound irreversibly inhibits the Bruton's tyrosine kinase to which itis covalently bound. In a further or alternative embodiment, thecompound forms a covalent bond with a cysteine residue on Bruton'styrosine kinase. In yet another aspect, provided herein is a method fortreating a cancer by administering to a subject in need thereof acomposition containing a therapeutically effective amount of a compoundthat forms a covalent bond with Bruton's tyrosine kinase. In oneembodiment, the compound forms a covalent bound with the activated formof Bruton's tyrosine kinase. In further or alternative embodiments, thecompound irreversibly inhibits the Bruton's tyrosine kinase to which itis covalently bound. In a further or alternative embodiment, thecompound forms a covalent bond with a cysteine residue on Bruton'styrosine kinase.

In another aspect, provided herein is a method for treating athromboembolic disorder by administering to a subject in need thereof acomposition containing a therapeutically effective amount of a compoundthat forms a covalent bond with Bruton's tyrosine kinase. In oneembodiment, the compound forms a covalent bound with the activated formof Bruton's tyrosine kinase. In further or alternative embodiments, thecompound irreversibly inhibits the Bruton's tyrosine kinase to which itis covalently bound. In a further or alternative embodiment, thecompound forms a covalent bond with a cysteine residue on Bruton'styrosine kinase.

In another aspect are methods for modulating, including irreversiblyinhibiting the activity of Btk or other tyrosine kinases, wherein theother tyrosine kinases share homology with Btk by having a cysteineresidue (including a Cys 481 residue) that can form a covalent bond withat least one irreversible inhibitor described herein, in a mammalcomprising administering to the mammal at least once an effective amountof at least one compound having the structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). In another aspect are methods for modulating, includingirreversibly inhibiting, the activity of Btk in a mammal comprisingadministering to the mammal at least once an effective amount of atleast one compound having the structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). In another aspect are methods for treating Btk-dependentor Btk mediated conditions or diseases, comprising administering to themammal at least once an effective amount of at least one compound havingthe structure of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh).

In another aspect are methods for treating inflammation comprisingadministering to the mammal at least once an effective amount of atleast one compound having the structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh).

A further aspect are methods for the treatment of cancer comprisingadministering to the mammal at least once an effective amount of atleast one compound having the structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). The type of cancer may include, but is not limited to,pancreatic cancer and other solid or hematological tumors.

In another aspect are methods for treating respiratory diseasescomprising administering to the mammal at least once an effective amountof at least one compound having the structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). In a further embodiment of this aspect, the respiratorydisease is asthma. In a further embodiment of this aspect, therespiratory disease includes, but is not limited to, adult respiratorydistress syndrome and allergic (extrinsic) asthma, non-allergic(intrinsic) asthma, acute severe asthma, chronic asthma, clinicalasthma, nocturnal asthma, allergen-induced asthma, aspirin-sensitiveasthma, exercise-induced asthma, isocapnic hyperventilation, child-onsetasthma, adult-onset asthma, cough-variant asthma, occupational asthma,steroid-resistant asthma, and seasonal asthma.

In another aspect are methods for preventing rheumatoid arthritis andosteoarthritis comprising administering to the mammal at least once aneffective amount of at least one compound having the structure ofFormula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh).

In another aspect are methods for treating inflammatory responses of theskin comprising administering to the mammal at least once an effectiveamount of at least one compound having the structure of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh). Such inflammatory responses of the skininclude, by way of example, dermatitis, contact dermatitis, eczema,urticaria, rosacea, and scarring. In another aspect are methods forreducing psoriatic lesions in the skin, joints, or other tissues ororgans, comprising administering to the mammal an effective amount of afirst compound having the structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh).

In another aspect is the use of a compound of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) in the manufacture of a medicament for treating aninflammatory disease or condition in an animal in which the activity ofBtk or other tyrosine kinases, wherein the other tyrosine kinases sharehomology with Btk by having a cysteine residue (including a Cys 481residue) that can form a covalent bond with at least one irreversibleinhibitor described herein, contributes to the pathology and/or symptomsof the disease or condition. In one embodiment of this aspect, thetyrosine kinase protein is Btk. In another or further embodiment of thisaspect, the inflammatory disease or conditions are respiratory,cardiovascular, or proliferative diseases.

In any of the aforementioned aspects are further embodiments in whichadministration is enteral, parenteral, or both, and wherein (a) theeffective amount of the compound is systemically administered to themammal; (b) the effective amount of the compound is administered orallyto the mammal; (c) the effective amount of the compound is intravenouslyadministered to the mammal; (d) the effective amount of the compoundadministered by inhalation; (e) the effective amount of the compound isadministered by nasal administration; or (f) the effective amount of thecompound is administered by injection to the mammal; (g) the effectiveamount of the compound is administered topically (dermal) to the mammal;(h) the effective amount of the compound is administered by ophthalmicadministration; or (i) the effective amount of the compound isadministered rectally to the mammal.

In any of the aforementioned aspects are further embodiments comprisingsingle administrations of the effective amount of the compound,including further embodiments in which (i) the compound is administeredonce; (ii) the compound is administered to the mammal multiple timesover the span of one day; (iii) continually; or (iv) continuously.

In any of the aforementioned aspects are further embodiments comprisingmultiple administrations of the effective amount of the compound,including further embodiments in which (i) the compound is administeredin a single dose; (ii) the time between multiple administrations isevery 6 hours; (iii) the compound is administered to the mammal every 8hours. In further or alternative embodiments, the method comprises adrug holiday, wherein the administration of the compound is temporarilysuspended or the dose of the compound being administered is temporarilyreduced; at the end of the drug holiday, dosing of the compound isresumed. The length of the drug holiday can vary from 2 days to 1 year.

In any of the aforementioned aspects involving the treatment ofproliferative disorders, including cancer, are further embodimentscomprising administering at least one additional agent selected from thegroup consisting of alemtuzumab, arsenic trioxide, asparaginase(pegylated or non-), bevacizumab, cetuximab, platinum-based compoundssuch as cisplatin, cladribine, daunorubicin/doxorubicin/idarubicin,irinotecan, fludarabine, 5-fluorouracil, gemtuzumab, methotrexate,Paclitaxel™, taxol, temozolomide, thioguanine, or classes of drugsincluding hormones (an antiestrogen, an antiandrogen, or gonadotropinreleasing hormone analogues, interferons such as alpha interferon,nitrogen mustards such as busulfan or melphalan or mechlorethamine,retinoids such as tretinoin, topoisomerase inhibitors such as irinotecanor topotecan, tyrosine kinase inhibitors such as gefinitinib orimatinib, or agents to treat signs or symptoms induced by such therapyincluding allopurinol, filgrastim, granisetron/ondansetron/palonosetron,dronabinol.

In any of the aforementioned aspects involving the prevention ortreatment of Btk-dependent or tyrosine kinase mediated diseases orconditions are further embodiments comprising identifying patients byscreening for a tyrosine kinase gene haplotype. In further oralternative embodiments the tyrosine kinase gene haplotype is a tyrosinekinase pathway gene, while in still further or alternative embodiments,the tyrosine kinase gene haplotype is a Btk haplotype.

In a further or alternative embodiment, the compounds of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) are irreversible inhibitors of Bruton'styrosine kinase (Btk), while in still further or alternativeembodiments, such irreversible inhibitors are selective for Btk. In evenfurther or alternative embodiments, such inhibitors have an IC₅₀ below10 microM in enzyme assay. In one embodiment, a Btk irreversibleinhibitor has an IC₅₀ of less than 1 microM, and in another embodiment,less than 0.25 microM.

In further or alternative embodiments, the compounds of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) are selective irreversible inhibitors forBtk over Itk. In further or alternative embodiments, the compounds ofFormula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) are selective irreversibleinhibitors for Btk over Lck. In further or alternative embodiments, thecompounds of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) are selectiveirreversible inhibitors for Btk over ABL. In further or alternativeembodiments, the compounds of (I), (IIa)-(IIb), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) are selectiveirreversible inhibitors for Btk over CMET. In further or alternativeembodiments, the compounds of Formula (I), (Ia)-(Ic), (IIa)-(IId),(IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId),(VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh)are selective irreversible inhibitors for Btk over EGFR. In further oralternative embodiments, the compounds of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) are selective irreversible inhibitors for Btk over Lyn.

In further or alternative embodiments, the irreversible Btk inhibitorsare also inhibitors of EGFR.

Other objects, features and advantages of the methods and compositionsdescribed herein will become apparent from the following detaileddescription. It should be understood, however, that the detaileddescription and the specific examples, while indicating specificembodiments, are given by way of illustration only, since variouschanges and modifications within the spirit and scope of the presentdisclosure will become apparent to those skilled in the art from thisdetailed description. The section headings used herein are fororganizational purposes only and are not to be construed as limiting thesubject matter described. All documents, or portions of documents, citedin the application including, but not limited to, patents, patentapplications, articles, books, manuals, and treatises are herebyexpressly incorporated by reference in their entirety for any purpose.

DETAILED DESCRIPTION OF THE INVENTION Certain Terminology

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart to which the claimed subject matter belongs. In the event that thereare a plurality of definitions for terms herein, those in this sectionprevail. Where reference is made to a URL or other such identifier oraddress, it is understood that such identifiers can change andparticular information on the internet can come and go, but equivalentinformation can be found by searching the internet. Reference theretoevidences the availability and public dissemination of such information.

It is to be understood that the foregoing general description and thefollowing detailed description are exemplary and explanatory only andare not restrictive of any subject matter claimed. In this application,the use of the singular includes the plural unless specifically statedotherwise. It must be noted that, as used in the specification and theappended claims, the singular forms “a,” “an” and “the” include pluralreferents unless the context clearly dictates otherwise. In thisapplication, the use of “or” means “and/or” unless stated otherwise.Furthermore, use of the term “including” as well as other forms, such as“include”, “includes,” and “included,” is not limiting.

Definition of standard chemistry terms may be found in reference works,including Carey and Sundberg “ADVANCED ORGANIC CHEMISTRY 4^(TH) ED.”Vols. A (2000) and B (2001), Plenum Press, New York. Unless otherwiseindicated, conventional methods of mass spectroscopy, NMR, HPLC, proteinchemistry, biochemistry, recombinant DNA techniques and pharmacology,within the skill of the art are employed. Unless specific definitionsare provided, the nomenclature employed in connection with, and thelaboratory procedures and techniques of, analytical chemistry, syntheticorganic chemistry, and medicinal and pharmaceutical chemistry describedherein are those known in the art. Standard techniques can be used forchemical syntheses, chemical analyses, pharmaceutical preparation,formulation, and delivery, and treatment of patients. Standardtechniques can be used for recombinant DNA, oligonucleotide synthesis,and tissue culture and transformation (e.g., electroporation,lipofection). Reactions and purification techniques can be performede.g., using kits of manufacturer's specifications or as commonlyaccomplished in the art or as described herein. The foregoing techniquesand procedures can be generally performed of conventional methods wellknown in the art and as described in various general and more specificreferences that are cited and discussed throughout the presentspecification.

It is to be understood that the methods and compositions describedherein are not limited to the particular methodology, protocols, celllines, constructs, and reagents described herein and as such may vary.It is also to be understood that the terminology used herein is for thepurpose of describing particular embodiments only, and is not intendedto limit the scope of the methods and compositions described herein,which will be limited only by the appended claims.

All publications and patents mentioned herein are incorporated herein byreference in their entirety for the purpose of describing anddisclosing, for example, the constructs and methodologies that aredescribed in the publications, which might be used in connection withthe methods, compositions and compounds described herein. Thepublications discussed herein are provided solely for their disclosureprior to the filing date of the present application. Nothing herein isto be construed as an admission that the inventors described herein arenot entitled to antedate such disclosure by virtue of prior invention orfor any other reason.

“Alkyl” refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, containing nounsaturation, having from one to fifteen carbon atoms (e.g., C₁-C₁₅alkyl). In certain embodiments, an alkyl comprises one to thirteencarbon atoms (e.g., C₁-C₁₃ alkyl). In certain embodiments, an alkylcomprises one to eight carbon atoms (e.g., C₁-C₈ alkyl). In otherembodiments, an alkyl comprises five to fifteen carbon atoms (e.g.,C₅-C₁₅ alkyl). In other embodiments, an alkyl comprises five to eightcarbon atoms (e.g., C₅-C₈ alkyl). The alkyl is attached to the rest ofthe molecule by a single bond, for example, methyl (Me), ethyl (Et),n-propyl (n-pr), 1-methylethyl (iso-propyl or i-Pr), n-butyl (n-Bu),n-pentyl, 1,1-dimethylethyl (t-butyl, or t-Bu), 3-methylhexyl,2-methylhexyl, and the like. Unless stated otherwise specifically in thespecification, an alkyl group is optionally substituted by one or moreof the following substituents: halo, cyano, nitro, oxo, thioxo,trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a),—N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2) and —S(O)_(t)N(R^(a))₂ (where t is 1or 2) where each R^(a) is independently hydrogen, alkyl, fluoroalkyl,carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl.

The alkyl group could also be a “lower alkyl” having 1 to 6 carbonatoms.

As used herein, C₁-C_(x) includes C₁-C₂, C₁-C₃ . . . C₁-C_(x).

“Alkenyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one double bond, and having from two to twelve carbon atoms. Incertain embodiments, an alkenyl comprises two to eight carbon atoms. Inother embodiments, an alkenyl comprises two to four carbon atoms. Thealkenyl is attached to the rest of the molecule by a single bond, forexample, ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl,pent-1-enyl, penta-1,4-dienyl, and the like. Unless stated otherwisespecifically in the specification, an alkenyl group is optionallysubstituted by one or more of the following substituents: halo, cyano,nitro, oxo, thioxo, trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a),—N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where tis 1 or 2), —S(O)_(t)OR^(a) (where t is 1 or 2) and —S(O)_(t)N(R^(a))₂(where t is 1 or 2) where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl.

“Alkynyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one triple bond, having from two to twelve carbon atoms. Incertain embodiments, an alkynyl comprises two to eight carbon atoms. Inother embodiments, an alkynyl has two to four carbon atoms. The alkynylis attached to the rest of the molecule by a single bond, for example,ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unlessstated otherwise specifically in the specification, an alkynyl group isoptionally substituted by one or more of the following substituents:halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, —OR^(a), —SR^(a),—OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where tis 1 or 2), —S(O)_(t)OR^(a) (where t is 1 or 2) and —S(O)_(t)N(R^(a))₂(where t is 1 or 2) where each R^(a) is independently hydrogen, alkyl,fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl.

“Alkylene” or “alkylene chain” refers to a straight or branched divalenthydrocarbon chain linking the rest of the molecule to a radical group,consisting solely of carbon and hydrogen, containing no unsaturation andhaving from one to twelve carbon atoms, for example, methylene,ethylene, propylene, n-butylene, and the like. The alkylene chain isattached to the rest of the molecule through a single bond and to theradical group through a single bond. The points of attachment of thealkylene chain to the rest of the molecule and to the radical group canbe through one carbon in the alkylene chain or through any two carbonswithin the chain. Unless stated otherwise specifically in thespecification, an alkylene chain is optionally substituted by one ormore of the following substituents: halo, cyano, nitro, aryl,cycloalkyl, heterocyclyl, heteroaryl, oxo, thioxo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2) and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl,heteroaryl or heteroarylalkyl.

“Alkenylene” or “alkenylene chain” refers to a straight or brancheddivalent hydrocarbon chain linking the rest of the molecule to a radicalgroup, consisting solely of carbon and hydrogen, containing at least onedouble bond and having from two to twelve carbon atoms, for example,ethenylene, propenylene, n-butenylene, and the like. The alkenylenechain is attached to the rest of the molecule through a double bond or asingle bond and to the radical group through a double bond or a singlebond. The points of attachment of the alkenylene chain to the rest ofthe molecule and to the radical group can be through one carbon or anytwo carbons within the chain. Unless stated otherwise specifically inthe specification, an alkenylene chain is optionally substituted by oneor more of the following substituents: halo, cyano, nitro, aryl,cycloalkyl, heterocyclyl, heteroaryl, oxo, thioxo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —N(R^(a))C(O)R^(a),—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2) and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl,aryl (optionally substituted with one or more halo groups), aralkyl,heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, andwhere each of the above substituents is unsubstituted unless otherwiseindicated.

“Aryl” refers to a radical derived from an aromatic monocyclic ormulticyclic hydrocarbon ring system by removing a hydrogen atom from aring carbon atom. The aromatic monocyclic or multicyclic hydrocarbonring system contains only hydrogen and carbon from six to eighteencarbon atoms, where at least one of the rings in the ring system isfully unsaturated, i.e., it contains a cyclic, delocalized (4n+2)π-electron system in accordance with the Hückel theory. Aryl groupsinclude, but are not limited to, groups such as phenyl (Ph), fluorenyl,and naphthyl. Unless stated otherwise specifically in the specification,the term “aryl” or the prefix “ar-” (such as in “aralkyl”) is meant toinclude aryl radicals optionally substituted by one or more substituentsindependently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl,cyano, nitro, optionally substituted aryl, optionally substitutedaralkyl, optionally substituted aralkenyl, optionally substitutedaralkynyl, optionally substituted carbocyclyl, optionally substitutedcarbocyclylalkyl, optionally substituted heterocyclyl, optionallysubstituted heterocyclylalkyl, optionally substituted heteroaryl,optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a),—R^(b)—C(O)OR^(a), —R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a)(where t is 1 or 2) and —R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2),where each R^(a) is independently hydrogen, alkyl, fluoroalkyl,cycloalkyl, cycloalkylalkyl, aryl (optionally substituted with one ormore halo groups), aralkyl, heterocyclyl, heterocyclylalkyl, heteroarylor heteroarylalkyl, each R^(b) is independently a direct bond or astraight or branched alkylene or alkenylene chain, and R^(c) is astraight or branched alkylene or alkenylene chain, and where each of theabove substituents is unsubstituted unless otherwise indicated.

“Aralkyl” refers to a radical of the formula —R^(c)-aryl where R^(c) isan alkylene chain as defined above, for example, benzyl, diphenylmethyland the like. The alkylene chain part of the aralkyl radical isoptionally substituted as described above for an alkylene chain. Thearyl part of the aralkyl radical is optionally substituted as describedabove for an aryl group.

“Aralkenyl” refers to a radical of the formula —R^(d)-aryl where R^(d)is an alkenylene chain as defined above. The aryl part of the aralkenylradical is optionally substituted as described above for an aryl group.The alkenylene chain part of the aralkenyl radical is optionallysubstituted as defined above for an alkenylene group.

“Aralkynyl” refers to a radical of the formula —R^(e)-aryl, where R^(e)is an alkynylene chain as defined above. The aryl part of the aralkynylradical is optionally substituted as described above for an aryl group.The alkynylene chain part of the aralkynyl radical is optionallysubstituted as defined above for an alkynylene chain.

“Carbocyclyl” refers to a stable non-aromatic monocyclic or polycyclichydrocarbon radical consisting solely of carbon and hydrogen atoms,which includes fused or bridged ring systems, having from three tofifteen carbon atoms. In certain embodiments, a carbocyclyl comprisesthree to ten carbon atoms. In other embodiments, a carbocyclyl comprisesfive to seven carbon atoms. The carbocyclyl is attached to the rest ofthe molecule by a single bond. Carbocyclyl is optionally saturated,(i.e., containing single C—C bonds only) or unsaturated (i.e.,containing one or more double bonds or triple bonds.) A fully saturatedcarbocyclyl radical is also referred to as “cycloalkyl.” Examples ofmonocyclic cycloalkyls include, e.g., cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. An unsaturatedcarbocyclyl is also referred to as “cycloalkenyl.” Examples ofmonocyclic cycloalkenyls include, e.g., cyclopentenyl, cyclohexenyl,cycloheptenyl, and cyclooctenyl. Polycyclic carbocyclyl radicalsinclude, for example, adamantyl, norbornyl (i.e.,bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl,7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwisestated specifically in the specification, the term “carbocyclyl” ismeant to include carbocyclyl radicals that are optionally substituted byone or more substituents independently selected from alkyl, alkenyl,alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionallysubstituted aryl, optionally substituted aralkyl, optionally substitutedaralkenyl, optionally substituted aralkynyl, optionally substitutedcarbocyclyl, optionally substituted carbocyclylalkyl, optionallysubstituted heterocyclyl, optionally substituted heterocyclylalkyl,optionally substituted heteroaryl, optionally substitutedheteroarylalkyl, —R^(b)—OR^(a), —R^(b)—SR^(a), —R^(b)—OC(O)—R^(a),—R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a),—R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a)(where t is 1 or 2) and —R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2),where each R^(a) is independently hydrogen, alkyl, fluoroalkyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl,heterocyclylalkyl, heteroaryl or heteroarylalkyl, each R^(b) isindependently a direct bond or a straight or branched alkylene oralkenylene chain, and R^(c) is a straight or branched alkylene oralkenylene chain, and where each of the above substituents isunsubstituted unless otherwise indicated.

“Halo” or “halogen” refers to bromo, chloro, fluoro or iodosubstituents.

The terms “haloalkyl,” “haloalkenyl,” “haloalkynyl” and “haloalkoxy”include alkyl, alkenyl, alkynyl and alkoxy structures in which at leastone hydrogen is replaced with a halogen atom. In certain embodiments inwhich two or more hydrogen atoms are replaced with halogen atoms, thehalogen atoms are all the same as one another. In other embodiments inwhich two or more hydrogen atoms are replaced with halogen atoms, thehalogen atoms are not all the same as one another.

“Fluoroalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more fluoro radicals, as defined above, forexample, trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl,1-fluoromethyl-2-fluoroethyl, and the like. The alkyl part of thefluoroalkyl radical is optionally substituted as defined above for analkyl group.

As used herein, the term “non-aromatic heterocycle”, “heterocycloalkyl”or “heteroalicyclic” refers to a non-aromatic ring wherein one or moreatoms forming the ring is a heteroatom. A “non-aromatic heterocycle” or“heterocycloalkyl” group refers to a cycloalkyl group that includes atleast one heteroatom selected from nitrogen, oxygen and sulfur. Theradicals may be fused with an aryl or heteroaryl. Heterocycloalkyl ringscan be formed by three to 14 ring atoms, such as three, four, five, six,seven, eight, nine, or more than nine atoms. Heterocycloalkyl rings canbe optionally substituted. In certain embodiments, non-aromaticheterocycles contain one or more carbonyl or thiocarbonyl groups suchas, for example, oxo- and thio-containing groups. Examples ofheterocycloalkyls include, but are not limited to, lactams, lactones,cyclic imides, cyclic thioimides, cyclic carbamates,tetrahydrothiopyran, 4H-pyran, tetrahydropyran, piperidine, 1,3-dioxin,1,3-dioxane, 1,4-dioxin, 1,4-dioxane, piperazine, 1,3-oxathiane,1,4-oxathiin, 1,4-oxathiane, tetrahydro-1,4-thiazine, 2H-1,2-oxazine,maleimide, succinimide, barbituric acid, thiobarbituric acid,dioxopiperazine, hydantoin, dihydrouracil, morpholine, trioxane,hexahydro-1,3,5-triazine, tetrahydrothiophene, tetrahydropyran,pyrroline, pyrrolidine, pyrrolidone, pyrrolidione, pyrazoline,pyrazolidine, imidazoline, imidazolidine, 1,3-dioxole, 1,3-dioxolane,1,3-dithiole, 1,3-dithiolane, isoxazoline, isoxazolidine, oxazoline,oxazolidine, oxazolidinone, thiazoline, thiazolidine, and1,3-oxathiolane. Illustrative examples of heterocycloalkyl groups, alsoreferred to as non-aromatic heterocycles, include:

and the like. The term heteroalicyclic also includes all ring forms ofthe carbohydrates, including but not limited to the monosaccharides, thedisaccharides and the oligosaccharides. Depending on the structure, aheterocycloalkyl group can be a monoradical or a diradical (i.e., aheterocycloalkylene group).

“Heteroaryl” refers to a radical derived from a 3- to 18-memberedaromatic ring radical that comprises two to seventeen carbon atoms andfrom one to six heteroatoms selected from nitrogen, oxygen and sulfur.As used herein, the heteroaryl radical is a monocyclic, bicyclic,tricyclic or tetracyclic ring system, wherein at least one of the ringsin the ring system is fully unsaturated, i.e., it contains a cyclic,delocalized (4n+2) π-electron system in accordance with the Hückeltheory. Heteroaryl includes fused or bridged ring systems. In someembodiments, heteroaryl rings have five, six, seven, eight, nine, ormore than nine ring atoms. The heteroatom(s) in the heteroaryl radicalis optionally oxidized. One or more nitrogen atoms, if present, areoptionally quaternized. The heteroaryl is attached to the rest of themolecule through any atom of the ring(s). Examples of heteroarylsinclude, but are not limited to, azepinyl, acridinyl, benzimidazolyl,benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl,benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl,benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl,benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl,benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl(benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,cyclopenta[d]pyrimidinyl,6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl,6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl,dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl,indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl,isoquinolyl, indolizinyl, isoxazolyl,5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl,1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl,5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl,phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl,purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl,pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl,pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl,quinolinyl, isoquinolinyl, tetrahydroquinolinyl,5,6,7,8-tetrahydroquinazolinyl,5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl,6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl,triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl,thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e.thienyl). Unless stated otherwise specifically in the specification, theterm “heteroaryl” is meant to include heteroaryl radicals as definedabove which are optionally substituted by one or more substituentsselected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl,haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl,optionally substituted aralkyl, optionally substituted aralkenyl,optionally substituted aralkynyl, optionally substituted carbocyclyl,optionally substituted carbocyclylalkyl, optionally substitutedheterocyclyl, optionally substituted heterocyclylalkyl, optionallysubstituted heteroaryl, optionally substituted heteroarylalkyl,—R^(b)—OR^(a), —R^(b)—SR^(a), —R^(b)—OC(O)—R^(a), —R^(b)—N(R^(a))₂,—R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a), —R^(b)—C(O)N(R^(a))₂,—R^(b)—O—R^(c)—C(O)N(R^(a))₂, —R^(b)—N(R^(a))C(O)OR^(a),—R^(b)—N(R^(a))C(O)R^(a), —R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or2), —R^(b)—S(O)_(t)OR^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl, fluoroalkyl, cycloalkyl, cycloalkylalkyl,aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl orheteroarylalkyl, each R^(b) is independently a direct bond or a straightor branched alkylene or alkenylene chain, and R^(c) is a straight orbranched alkylene or alkenylene chain, and where each of the abovesubstituents is unsubstituted unless otherwise indicated.

“N-heteroaryl” refers to a heteroaryl radical as defined abovecontaining at least one nitrogen and where the point of attachment ofthe heteroaryl radical to the rest of the molecule is through a nitrogenatom in the heteroaryl radical. An N-heteroaryl radical is optionallysubstituted as described above for heteroaryl radicals.

“C-heteroaryl” refers to a heteroaryl radical as defined above and wherethe point of attachment of the heteroaryl radical to the rest of themolecule is through a carbon atom in the heteroaryl radical. AC-heteroaryl radical is optionally substituted as described above forheteroaryl radicals.

“Heteroarylalkyl” refers to a radical of the formula —R^(c)-heteroaryl,where R^(c) is an alkylene chain as defined above. If the heteroaryl isa nitrogen-containing heteroaryl, the heteroaryl is optionally attachedto the alkyl radical at the nitrogen atom. The alkylene chain of theheteroarylalkyl radical is optionally substituted as defined above foran alkylene chain. The heteroaryl part of the heteroarylalkyl radical isoptionally substituted as defined above for a heteroaryl group.

“Sulfanyl” refers to the —S— radical.

“Sulfinyl” refers to the —S(═O)— radical.

“Sulfonyl” refers to the —S(═O)₂— radical.

“Amino” refers to the —NH₂ radical.

“Cyano” refers to the —CN radical.

“Nitro” refers to the —NO₂ radical.

“Oxa” refers to the —O— radical.

“Oxo” refers to the ═O radical.

“Imino” refers to the ═NH radical.

“Thioxo” refers to the ═S radical.

An “alkoxy” group refers to a (alkyl)O— group, where alkyl is as definedherein.

An “aryloxy” group refers to an (aryl)O— group, where aryl is as definedherein.

“Carbocyclylalkyl” means an alkyl radical, as defined herein,substituted with a carbocyclyl group. “Cycloalkylalkyl” means an alkylradical, as defined herein, substituted with a cycloalkyl group.Non-limiting cycloalkylalkyl groups include cyclopropylmethyl,cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, and the like.

As used herein, the terms “heteroalkyl” “heteroalkenyl” and“heteroalkynyl” include optionally substituted alkyl, alkenyl andalkynyl radicals in which one or more skeletal chain atoms is aheteroatom, e.g., oxygen, nitrogen, sulfur, silicon, phosphorus orcombinations thereof. The heteroatom(s) may be placed at any interiorposition of the heteroalkyl group or at the position at which theheteroalkyl group is attached to the remainder of the molecule. Examplesinclude, but are not limited to, —CH₂—O—CH₃, —CH₂—CH₂—O—CH₃,—CH₂—NH—CH₃, —CH₂—CH₂—NH—CH₃, —CH₂—N(CH₃)—CH₃, —CH₂—CH₂—NH—CH₃,—CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃, —CH₂—CH₂, —S(O)—CH₃,—CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —Si(CH₃)₃, —CH₂—CH═N—OCH₃, and—CH═CH—N(CH₃)—CH₃. In addition, up to two heteroatoms may beconsecutive, such as, by way of example, —CH₂—NH—OCH₃ and—CH₂—O—Si(CH₃)₃.

The term “heteroatom” refers to an atom other than carbon or hydrogen.Heteroatoms are typically independently selected from among oxygen,sulfur, nitrogen, silicon and phosphorus, but are not limited to theseatoms. In embodiments in which two or more heteroatoms are present, thetwo or more heteroatoms can all be the same as one another, or some orall of the two or more heteroatoms can each be different from theothers.

The term “bond,” “direct bond” or “single bond” refers to a chemicalbond between two atoms, or two moieties when the atoms joined by thebond are considered to be part of larger substructure.

An “isocyanato” group refers to a —NCO group.

An “isothiocyanato” group refers to a —NCS group.

The term “moiety” refers to a specific segment or functional group of amolecule. Chemical moieties are often recognized chemical entitiesembedded in or appended to a molecule.

A “thioalkoxy” or “alkylthio” group refers to a —S-alkyl group.

A “alkylthioalkyl” group refers to an alkyl group substituted with a—S-alkyl group.

As used herein, the term “acyloxy” refers to a group of formulaRC(═O)O—.

“Carboxy” means a —C(O)OH radical.

As used herein, the term “acetyl” refers to a group of formula—C(═O)CH₃.

“Acyl” refers to the group —C(O)R.

As used herein, the term “trihalomethanesulfonyl” refers to a group offormula X₃CS(═O)₂— where X is a halogen.

“Cyanoalkyl” means an alkyl radical, as defined herein, substituted withat least one cyano group.

As used herein, the term “N-sulfonamido” or “sulfonylamino” refers to agroup of formula RS(═O)₂NH—.

As used herein, the term “O-carbamyl” refers to a group of formula—OC(═O)NR₂.

As used herein, the term “N-carbamyl” refers to a group of formulaROC(═O)NH—.

As used herein, the term “O-thiocarbamyl” refers to a group of formula—OC(═S)NR₂.

As used herein, “N-thiocarbamyl” refers to a group of formulaROC(═S)NH—.

As used herein, the term “C-amido” refers to a group of formula—C(═O)NR₂.

“Aminocarbonyl” refers to a —CONH₂ radical.

As used herein, the term “N-amido” refers to a group of formulaRC(═O)NH—.

As used herein, the substituent “R” appearing by itself and without anumber designation refers to a substituent selected from among fromalkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) andnon-aromatic heterocycle (bonded through a ring carbon).

“Hydroxyalkyl” refers to an alkyl radical, as defined herein,substituted with at least one hydroxy group. Non-limiting examples of ahydroxyalkyl include, but are not limited to, hydroxymethyl,2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl,1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl,4-hydroxybutyl, 2,3-dihydroxypropyl, 1-(hydroxymethyl)-2-hydroxyethyl,2,3-dihydroxybutyl, 3,4-dihydroxybutyl and2-(hydroxymethyl)-3-hydroxypropyl.

“Alkoxyalkyl” refers to an alkyl radical, as defined herein, substitutedwith an alkoxy group, as defined herein.

An “alkenyloxy” group refers to a (alkenyl)O— group, where alkenyl is asdefined herein.

The term “alkylamine” refers to the —N(alkyl)_(x)H_(y) group, where xand y are selected from among x=1, y=1 and x=2, y=0. When x=2, the alkylgroups, taken together with the N atom to which they are attached, canoptionally form a cyclic ring system.

“Alkylaminoalkyl” refers to an alkyl radical, as defined herein,substituted with an alkylamine, as defined herein.

An “amide” is a chemical moiety with the formula —C(O)NHR or —NHC(O)R,where R is selected from among alkyl, cycloalkyl, aryl, heteroaryl(bonded through a ring carbon) and heteroalicyclic (bonded through aring carbon). An amide moiety may form a linkage between an amino acidor a peptide molecule and a compound described herein, thereby forming aprodrug. Any amine, or carboxyl side chain on the compounds describedherein can be amidified. The procedures and specific groups to make suchamides are known to those of skill in the art and can readily be foundin reference sources such as Greene and Wuts, Protective Groups inOrganic Synthesis, 3^(rd) Ed., John Wiley & Sons, New York, N.Y., 1999,which is incorporated herein by reference in its entirety.

The term “ester” refers to a chemical moiety with formula —COOR, where Ris selected from among alkyl, cycloalkyl, aryl, heteroaryl (bondedthrough a ring carbon) and heteroalicyclic (bonded through a ringcarbon). Any hydroxy, or carboxyl side chain on the compounds describedherein can be esterified. The procedures and specific groups to makesuch esters are known to those of skill in the art and can readily befound in reference sources such as Greene and Wuts, Protective Groups inOrganic Synthesis, 3^(rd) Ed., John Wiley & Sons, New York, N.Y., 1999,which is incorporated herein by reference in its entirety.

As used herein, the term “ring” refers to any covalently closedstructure. Rings include, for example, carbocycles (e.g., aryls andcycloalkyls), heterocycles (e.g., heteroaryls and non-aromaticheterocycles), aromatics (e.g. aryls and heteroaryls), and non-aromatics(e.g., cycloalkyls and non-aromatic heterocycles). Rings can beoptionally substituted. Rings can be monocyclic or polycyclic.

As used herein, the term “ring system” refers to one, or more than onering.

The term “membered ring” can embrace any cyclic structure. The term“membered” is meant to denote the number of skeletal atoms thatconstitute the ring. Thus, for example, cyclohexyl, pyridine, pyran andthiopyran are 6-membered rings and cyclopentyl, pyrrole, furan, andthiophene are 5-membered rings.

The term “fused” refers to structures in which two or more rings shareone or more bonds.

The term “optionally substituted” or “substituted” means that thereferenced group may be substituted with one or more additional group(s)individually and independently selected from alkyl, cycloalkyl, aryl,heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, alkylthio,arylthio, alkylsulfoxide, arylsulfoxide, alkylsulfone, arylsulfone,cyano, halo, acyl, nitro, haloalkyl, fluoroalkyl, amino, including mono-and di-substituted amino groups, and the protected derivatives thereof.By way of example an optional substituents may be L^(s)R^(s), whereineach L^(s) is independently selected from a bond, —O—, —C(═O)—, —S—,—S(═O)—, —S(═O)₂—, —NH—, —NHC(O)—, —C(O)NH—, S(═O)₂NH—, —NHS(═O)₂,—OC(O)NH—, —NHC(O)O—, -(substituted or unsubstituted C₁-C₆ alkyl), or-(substituted or unsubstituted C₂-C₆ alkenyl); and each R^(s) isindependently selected from H, (substituted or unsubstitutedC₁-C₄alkyl), (substituted or unsubstituted C₃-C₆cycloalkyl), aryl,heteroaryl, or heteroalkyl. The protecting groups that may form theprotective derivatives of the above substituents are known to those ofskill in the art and may be found in references such as Greene and Wuts,above.

The term “nucleophile” or “nucleophilic” refers to an electron richcompound, or moiety thereof. An example of a nucleophile includes, butin no way is limited to, a cysteine residue of a molecule, such as, forexample Cys 481 of Btk.

The term “electrophile”, or “electrophilic” refers to an electron pooror electron deficient molecule, or moiety thereof. Examples ofelectrophiles include, but in no way are limited to, Michael acceptormoieties.

The term “acceptable” or “pharmaceutically acceptable”, with respect toa formulation, composition or ingredient, as used herein, means havingno persistent detrimental effect on the general health of the subjectbeing treated or does not abrogate the biological activity or propertiesof the compound, and is relatively nontoxic.

As used herein, the term “agonist” refers to a compound, the presence ofwhich results in a biological activity of a protein that is the same asthe biological activity resulting from the presence of a naturallyoccurring ligand for the protein, such as, for example, Btk.

As used herein, the term “partial agonist” refers to a compound thepresence of which results in a biological activity of a protein that isof the same type as that resulting from the presence of a naturallyoccurring ligand for the protein, but of a lower magnitude.

As used herein, the term “antagonist” refers to a compound, the presenceof which results in a decrease in the magnitude of a biological activityof a protein. In certain embodiments, the presence of an antagonistresults in complete inhibition of a biological activity of a protein,such as, for example, Btk. In certain embodiments, an antagonist is aninhibitor.

As used herein, “amelioration” of the symptoms of a particular disease,disorder or condition by administration of a particular compound orpharmaceutical composition refers to any lessening of severity, delay inonset, slowing of progression, or shortening of duration, whetherpermanent or temporary, lasting or transient that can be attributed toor associated with administration of the compound or composition.

“Bioavailability” refers to the percentage of the weight of compoundsdisclosed herein, such as, compounds of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) dosed that isdelivered into the general circulation of the animal or human beingstudied. The total exposure (AUC_((0-∞))) of a drug when administeredintravenously is usually defined as 100% bioavailable (F %). “Oralbioavailability” refers to the extent to which compounds disclosedherein, such as, compounds of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) areabsorbed into the general circulation when the pharmaceuticalcomposition is taken orally as compared to intravenous injection.

“Blood plasma concentration” refers to the concentration of compoundsdisclosed herein, such as, compounds of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) in the plasma component of blood of a subject. It isunderstood that the plasma concentration of compounds of any of Formula(I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) may vary significantly between subjects,due to variability with respect to metabolism and/or possibleinteractions with other therapeutic agents. In accordance with oneembodiment disclosed herein, the blood plasma concentration of thecompounds of any of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) may vary fromsubject to subject. Likewise, values such as maximum plasmaconcentration (C_(max)) or time to reach maximum plasma concentration(T_(max)), or total area under the plasma concentration time curve(AUC_((0-∞))) may vary from subject to subject. Due to this variability,the amount necessary to constitute “a therapeutically effective amount”of a compound of any of Formula (I), (Ia)-(Ic), (IIa)-(IId),(IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId),(VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh)may vary from subject to subject.

The term “Bruton's tyrosine kinase,” as used herein, refers to Bruton'styrosine kinase from Homo sapiens, as disclosed in, e.g., U.S. Pat. No.6,326,469 (GenBank Accession No. NP_(—)000052).

The term “Bruton's tyrosine kinase homolog,” as used herein, refers toorthologs of Bruton's tyrosine kinase, e.g., the orthologs from mouse(GenBank Acession No. AAB47246), dog (GenBank Accession No.XP_(—)549139.), rat (GenBank Acession No. NP_(—)001007799), chicken(GenBank Acession No. NP_(—)989564), or zebra fish (GenBank Acession No.XP_(—)698117), and fusion proteins of any of the foregoing that exhibitkinase activity towards one or more substrates of Bruton's tyrosinekinase (e.g. a peptide substrate having the amino acid sequence“AVLESEEELYSSARQ”).

The terms “co-administration” or the like, as used herein, are meant toencompass administration of the selected therapeutic agents to a singlepatient, and are intended to include treatment regimens in which theagents are administered by the same or different route of administrationor at the same or different time.

The terms “effective amount” or “therapeutically effective amount,” asused herein, refer to a sufficient amount of an agent or a compoundbeing administered which will relieve to some extent one or more of thesymptoms of the disease or condition being treated. The result can bereduction and/or alleviation of the signs, symptoms, or causes of adisease, or any other desired alteration of a biological system. Forexample, an “effective amount” for therapeutic uses is the amount of thecomposition including a compound as disclosed herein required to providea clinically significant decrease in disease symptoms without undueadverse side effects. An appropriate “effective amount” in anyindividual case may be determined using techniques, such as a doseescalation study. The term “therapeutically effective amount” includes,for example, a prophylactically effective amount. An “effective amount”of a compound disclosed herein is an amount effective to achieve adesired pharmacologic effect or therapeutic improvement without undueadverse side effects. It is understood that “an effect amount” or “atherapeutically effective amount” can vary from subject to subject, dueto variation in metabolism of the compound of any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh), age, weight, general condition of thesubject, the condition being treated, the severity of the conditionbeing treated, and the judgment of the prescribing physician. By way ofexample only, therapeutically effective amounts may be determined byroutine experimentation, including but not limited to a dose escalationclinical trial.

The terms “enhance” or “enhancing” means to increase or prolong eitherin potency or duration a desired effect. By way of example, “enhancing”the effect of therapeutic agents refers to the ability to increase orprolong, either in potency or duration, the effect of therapeutic agentson during treatment of a disease, disorder or condition. An“enhancing-effective amount,” as used herein, refers to an amountadequate to enhance the effect of a therapeutic agent in the treatmentof a disease, disorder or condition. When used in a patient, amountseffective for this use will depend on the severity and course of thedisease, disorder or condition, previous therapy, the patient's healthstatus and response to the drugs, and the judgment of the treatingphysician.

The term “homologous cysteine,” as used herein refers to a cysteineresidue found with in a sequence position that is homologous to that ofcysteine 481 of Bruton's tyrosine kinase, as defined herein. Forexample, cysteine 482 is the homologous cysteine of the rat ortholog ofBruton's tyrosine kinase; cysteine 479 is the homologous cysteine of thechicken ortholog; and cysteine 481 is the homologous cysteine in thezebra fish ortholog. In another example, the homologous cysteine of TXK,a Tec kinase family member related to Bruton's tyrosine, is Cys 350.

The term “identical,” as used herein, refers to two or more sequences orsubsequences which are the same. In addition, the term “substantiallyidentical,” as used herein, refers to two or more sequences which have apercentage of sequential units which are the same when compared andaligned for maximum correspondence over a comparison window, ordesignated region as measured using comparison algorithms or by manualalignment and visual inspection. By way of example only, two or moresequences may be “substantially identical” if the sequential units areabout 60% identical, about 65% identical, about 70% identical, about 75%identical, about 80% identical, about 85% identical, about 90%identical, or about 95% identical over a specified region. Suchpercentages to describe the “percent identity” of two or more sequences.The identity of a sequence can exist over a region that is at leastabout 75-100 sequential units in length, over a region that is about 50sequential units in length, or, where not specified, across the entiresequence. This definition also refers to the complement of a testsequence. By way of example only, two or more polypeptide sequences areidentical when the amino acid residues are the same, while two or morepolypeptide sequences are “substantially identical” if the amino acidresidues are about 60% identical, about 65% identical, about 70%identical, about 75% identical, about 80% identical, about 85%identical, about 90% identical, or about 95% identical over a specifiedregion. The identity can exist over a region that is at least about75-100 amino acids in length, over a region that is about 50 amino acidsin length, or, where not specified, across the entire sequence of apolypeptide sequence. In addition, by way of example only, two or morepolynucleotide sequences are identical when the nucleic acid residuesare the same, while two or more polynucleotide sequences are“substantially identical” if the nucleic acid residues are about 60%identical, about 65% identical, about 70% identical, about 75%identical, about 80% identical, about 85% identical, about 90%identical, or about 95% identical over a specified region. The identitycan exist over a region that is at least about 75-100 nucleic acids inlength, over a region that is about 50 nucleic acids in length, or,where not specified, across the entire sequence of a polynucleotidesequence.

The terms “inhibits”, “inhibiting”, or “inhibitor” of a kinase, as usedherein, refer to inhibition of enzymatic phosphotransferase activity.

The term “irreversible inhibitor,” as used herein, refers to a compoundthat, upon contact with a target protein (e.g., a kinase) causes theformation of a new covalent bond with or within the protein, whereby oneor more of the target protein's biological activities (e.g.,phosphotransferase activity) is diminished or abolished notwithstandingthe subsequent presence or absence of the irreversible inhibitor. Incontrast, a reversible inhibitor compound upon contact with a targetprotein does not cause the formation of a new covalent bond with orwithin the protein and therefore can associate and dissociate from thetarget protein.

The term “irreversible Btk inhibitor,” as used herein, refers to aninhibitor of Btk that can form a covalent bond with an amino acidresidue of Btk. In one embodiment, the irreversible inhibitor of Btk canform a covalent bond with a Cys residue of Btk; in particularembodiments, the irreversible inhibitor can form a covalent bond with aCys 481 residue (or a homolog thereof) of Btk or a cysteine residue inthe homologous corresponding position of another tyrosine kinase.

The term “isolated,” as used herein, refers to separating and removing acomponent of interest from components not of interest. Isolatedsubstances can be in either a dry or semi-dry state, or in solution,including but not limited to an aqueous solution. The isolated componentcan be in a homogeneous state or the isolated component can be a part ofa pharmaceutical composition that comprises additional pharmaceuticallyacceptable carriers and/or excipients. By way of example only, nucleicacids or proteins are “isolated” when such nucleic acids or proteins arefree of at least some of the cellular components with which it isassociated in the natural state, or that the nucleic acid or protein hasbeen concentrated to a level greater than the concentration of its invivo or in vitro production. Also, by way of example, a gene is isolatedwhen separated from open reading frames which flank the gene and encodea protein other than the gene of interest.

A “metabolite” of a compound disclosed herein is a derivative of thatcompound that is formed when the compound is metabolized. The term“active metabolite” refers to a biologically active derivative of acompound that is formed when the compound is metabolized. The term“metabolized,” as used herein, refers to the sum of the processes(including, but not limited to, hydrolysis reactions and reactionscatalyzed by enzymes, such as, oxidation reactions) by which aparticular substance is changed by an organism. Thus, enzymes mayproduce specific structural alterations to a compound. For example,cytochrome P450 catalyzes a variety of oxidative and reductive reactionswhile uridine diphosphate glucuronyl transferases catalyze the transferof an activated glucuronic-acid molecule to aromatic alcohols, aliphaticalcohols, carboxylic acids, amines and free sulfhydryl groups. Furtherinformation on metabolism may be obtained from The Pharmacological Basisof Therapeutics, 9th Edition, McGraw-Hill (1996). Metabolites of thecompounds disclosed herein can be identified either by administration ofcompounds to a host and analysis of tissue samples from the host, or byincubation of compounds with hepatic cells in vitro and analysis of theresulting compounds. Both methods are well known in the art. In someembodiments, metabolites of a compound are formed by oxidative processesand correspond to the corresponding hydroxy-containing compound. In someembodiments, a compound is metabolized to pharmacologically activemetabolites.

The term “modulate,” as used herein, means to interact with a targeteither directly or indirectly so as to alter the activity of the target,including, by way of example only, to enhance the activity of thetarget, to inhibit the activity of the target, to limit the activity ofthe target, or to extend the activity of the target.

As used herein, the term “modulator” refers to a compound that alters anactivity of a molecule. For example, a modulator can cause an increaseor decrease in the magnitude of a certain activity of a moleculecompared to the magnitude of the activity in the absence of themodulator. In certain embodiments, a modulator is an inhibitor, whichdecreases the magnitude of one or more activities of a molecule. Incertain embodiments, an inhibitor completely prevents one or moreactivities of a molecule. In certain embodiments, a modulator is anactivator, which increases the magnitude of at least one activity of amolecule. In certain embodiments the presence of a modulator results inan activity that does not occur in the absence of the modulator.

The term “prophylactically effective amount,” as used herein, refersthat amount of a composition applied to a patient that will relieve tosome extent one or more of the symptoms of a disease, condition ordisorder being treated. In such prophylactic applications, such amountsmay depend on the patient's state of health, weight, and the like. It isconsidered well within the skill of the art for one to determine suchprophylactically effective amounts by routine experimentation,including, but not limited to, a dose escalation clinical trial.

As used herein, the term “selective binding compound” refers to acompound that selectively binds to any portion of one or more targetproteins.

As used herein, the term “selectively binds” refers to the ability of aselective binding compound to bind to a target protein, such as, forexample, Btk, with greater affinity than it binds to a non-targetprotein. In certain embodiments, specific binding refers to binding to atarget with an affinity that is at least 10, 50, 100, 250, 500, 1000 ormore times greater than the affinity for a non-target.

As used herein, the term “selective modulator” refers to a compound thatselectively modulates a target activity relative to a non-targetactivity. In certain embodiments, specific modulator refers tomodulating a target activity at least 10, 50, 100, 250, 500, 1000 timesmore than a non-target activity.

The term “substantially purified,” as used herein, refers to a componentof interest that may be substantially or essentially free of othercomponents which normally accompany or interact with the component ofinterest prior to purification. By way of example only, a component ofinterest may be “substantially purified” when the preparation of thecomponent of interest contains less than about 30%, less than about 25%,less than about 20%, less than about 15%, less than about 10%, less thanabout 5%, less than about 4%, less than about 3%, less than about 2%, orless than about 1% (by dry weight) of contaminating components. Thus, a“substantially purified” component of interest may have a purity levelof about 70%, about 75%, about 80%, about 85%, about 90%, about 95%,about 96%, about 97%, about 98%, about 99% or greater.

The term “subject” or “patient” as used herein, refers to an animalwhich is the object of treatment, observation or experiment. By way ofexample only, a subject may be, but is not limited to, a mammalincluding, but not limited to, a human.

As used herein, the term “target activity” refers to a biologicalactivity capable of being modulated by a selective modulator. Certainexemplary target activities include, but are not limited to, bindingaffinity, signal transduction, enzymatic activity, tumor growth,inflammation or inflammation-related processes, and amelioration of oneor more symptoms associated with a disease or condition.

As used herein, the term “target protein” refers to a molecule or aportion of a protein capable of being bound by a selective bindingcompound. In certain embodiments, a target protein is Btk.

The terms “treat,” “treating” or “treatment”, as used herein, includealleviating, abating or ameliorating a disease or condition symptoms,preventing additional symptoms, ameliorating or preventing theunderlying metabolic causes of symptoms, inhibiting the disease orcondition, e.g., arresting the development of the disease or condition,relieving the disease or condition, causing regression of the disease orcondition, relieving a condition caused by the disease or condition, orstopping the symptoms of the disease or condition. The terms “treat,”“treating” or “treatment”, include, but are not limited to, prophylacticand/or therapeutic treatments.

As used herein, the IC₅₀ refers to an amount, concentration or dosage ofa particular test compound that achieves a 50% inhibition of a maximalresponse, such as inhibition of Btk, in an assay that measures suchresponse.

As used herein, EC₅₀ refers to a dosage, concentration or amount of aparticular test compound that elicits a dose-dependent response at 50%of maximal expression of a particular response that is induced, provokedor potentiated by the particular test compound.

The methods described herein include administering to a subject in needa composition containing a therapeutically effective amount of one ormore irreversible Btk inhibitor compounds described herein. Withoutbeing bound by theory, the diverse roles played by Btk signaling invarious hematopoietic cell functions, e.g., B-cell receptor activation,suggests that small molecule Btk inhibitors are useful for reducing therisk of or treating a variety of diseases affected by or affecting manycell types of the hematopoetic lineage including, e.g., autoimmunediseases, heteroimmune conditions or diseases, inflammatory diseases,cancer (e.g., B-cell proliferative disorders), and thromboembolicdisorders. Further, the irreversible Btk inhibitor compounds describedherein can be used to inhibit a small subset of other tyrosine kinasesthat share homology with Btk by having a cysteine residue (including aCys 481 residue) that can form a covalent bond with the irreversibleinhibitor. Thus, a subset of tyrosine kinases other than Btk are alsoexpected to be useful as therapeutic targets in a number of healthconditions.

In some embodiments, the methods described herein can be used to treatan autoimmune disease, which includes, but is not limited to, rheumatoidarthritis, psoriatic arthritis, osteoarthritis, Still's disease,juvenile arthritis, lupus, diabetes, myasthenia gravis, Hashimoto'sthyroiditis, Ord's thyroiditis, Graves' disease Sjögren's syndrome,multiple sclerosis, Guillain-Barré syndrome, acute disseminatedencephalomyelitis, Addison's disease, opsoclonus-myoclonus syndrome,ankylosing spondylitis, antiphospholipid antibody syndrome, aplasticanemia, autoimmune hepatitis, coeliac disease, Goodpasture's syndrome,idiopathic thrombocytopenic purpura, optic neuritis, scleroderma,primary biliary cirrhosis, Reiter's syndrome, Takayasu's arteritis,temporal arteritis, warm autoimmune hemolytic anemia, Wegener'sgranulomatosis, psoriasis, alopecia universalis, Behçet's disease,chronic fatigue, dysautonomia, endometriosis, interstitial cystitis,neuromyotonia, scleroderma, and vulvodynia.

In some embodiments, the methods described herein can be used to treatheteroimmune conditions or diseases, which include, but are not limitedto graft versus host disease, transplantation, transfusion, anaphylaxis,allergies (e.g., allergies to plant pollens, latex, drugs, foods, insectpoisons, animal hair, animal dander, dust mites, or cockroach calyx),type I hypersensitivity, allergic conjunctivitis, allergic rhinitis, andatopic dermatitis.

In further embodiments, the methods described herein can be used totreat an inflammatory disease, which includes, but is not limited toasthma, inflammatory bowel disease, appendicitis, blepharitis,bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis,cholecystitis, colitis, conjunctivitis, cystitis, dacryoadenitis,dermatitis, dermatomyositis, encephalitis, endocarditis, endometritis,enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis,fibrositis, gastritis, gastroenteritis, hepatitis, hidradenitissuppurativa, laryngitis, mastitis, meningitis, myelitis myocarditis,myositis, nephritis, oophoritis, orchitis, osteitis, otitis,pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis,pleuritis, phlebitis, pneumonitis, pneumonia, proctitis, prostatitis,pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis,tendonitis, tonsillitis, uveitis, vaginitis, vasculitis, and vulvitis.

In yet other embodiments, the methods described herein can be used totreat a cancer, e.g., B-cell proliferative disorders, which include, butare not limited to diffuse large B cell lymphoma, follicular lymphoma,chronic lymphocytic lymphoma, chronic lymphocytic leukemia, B-cellprolymphocytic leukemia, lymphoplasmacytic lymphoma/Waldenströmmacroglobulinemia, splenic marginal zone lymphoma, plasma cell myeloma,plasmacytoma, extranodal marginal zone B cell lymphoma, nodal marginalzone B cell lymphoma, mantle cell lymphoma, mediastinal (thymic) large Bcell lymphoma, intravascular large B cell lymphoma, primary effusionlymphoma, burkitt lymphoma/leukemia, and lymphomatoid granulomatosis.

In further embodiments, the methods described herein can be used totreat thromboembolic disorders, which include, but are not limited tomyocardial infarct, angina pectoris (including unstable angina),reocclusions or restenoses after angioplasty or aortocoronary bypass,stroke, transitory ischemia, peripheral arterial occlusive disorders,pulmonary embolisms, and deep venous thromboses.

Symptoms, diagnostic tests, and prognostic tests for each of theabove-mentioned conditions are known in the art. See, e.g., Harrison'sPrinciples of Internal Medicine©,” 16th ed., 2004, The McGraw-HillCompanies, Inc. Dey et al. (2006), Cytojournal 3(24), and the “RevisedEuropean American Lymphoma” (REAL) classification system (see, e.g., thewebsite maintained by the National Cancer Institute).

A number of animal models of are useful for establishing a range oftherapeutically effective doses of irreversible Btk inhibitor compoundsfor treating any of the foregoing diseases.

For example, dosing of irreversible Btk inhibitor compounds for treatingan autoimmune disease can be assessed in a mouse model of rheumatoidarthritis. In this model, arthritis is induced in Balb/c mice byadministering anti-collagen antibodies and lipopolysaccharide. SeeNandakumar et al. (2003), Am. J. Pathol 163:1827-1837.

In another example, dosing of irreversible Btk inhibitors for thetreatment of B-cell proliferative disorders can be examined in, e.g., ahuman-to-mouse xenograft model in which human B-cell lymphoma cells(e.g. Ramos cells) are implanted into immunodeficient mice (e.g., “nude”mice) as described in, e.g., Pagel et al. (2005), Clin Cancer Res11(13):4857-4866.

Animal models for treatment of thromboembolic disorders are also known.

The therapeutic efficacy of the compound for one of the foregoingdiseases can be optimized during a course of treatment. For example, asubject being treated can undergo a diagnostic evaluation to correlatethe relief of disease symptoms or pathologies to inhibition of in vivoBtk activity achieved by administering a given dose of an irreversibleBtk inhibitor. Cellular assays known in the art can be used to determinein vivo activity of Btk in the presence or absence of an irreversibleBtk inhibitor. For example, since activated Btk is phosphorylated attyrosine 223 (Y223) and tyrosine 551 (Y551), phospho-specificimmunocytochemical staining of P-Y223 or P-Y551-positive cells can beused to detect or quantify activation of Bkt in a population of cells(e.g., by FACS analysis of stained vs unstained cells). See, e.g.,Nisitani et al. (1999), Proc. Natl. Acad. Sci, USA 96:2221-2226. Thus,the amount of the Btk inhibitor compound that is administered to asubject can be increased or decreased as needed so as to maintain alevel of Btk inhibition optimal for treating the subject's diseasestate.

Compounds

In the following description of irreversible Btk compounds suitable foruse in the methods described herein, definitions of referred-to standardchemistry terms may be found in reference works (if not otherwisedefined herein), including Carey and Sundberg “Advanced OrganicChemistry 4th Ed.” Vols. A (2000) and B (2001), Plenum Press, New York.Unless otherwise indicated, conventional methods of mass spectroscopy,NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniquesand pharmacology, within the ordinary skill of the art are employed. Inaddition, nucleic acid and amino acid sequences for Btk (e.g., humanBtk) are known in the art as disclosed in, e.g., U.S. Pat. No.6,326,469. Unless specific definitions are provided, the nomenclatureemployed in connection with, and the laboratory procedures andtechniques of, analytical chemistry, synthetic organic chemistry, andmedicinal and pharmaceutical chemistry described herein are those knownin the art. Standard techniques can be used for chemical syntheses,chemical analyses, pharmaceutical preparation, formulation, anddelivery, and treatment of patients.

Further described are irreversible inhibitors of Btk that form acovalent bond with a serine residue on C481S mutated Btk. Specificallydescribed are irreversible inhibitors of Btk that form a covalent bondwith a serine481 residue on C481S mutated Btk (Woyach, et al. Resistancemechanisms for the Bruton's tyrosine kinase inhibitor ibrutinib, N EnglJ Med. 2014, 12; 370(24):2286-94). Further described herein areirreversible inhibitors of other tyrosine kinases, wherein the othertyrosine kinases share homology with C481S mutated Btk by having aserine residue (including a homologous residue to BTK C481S residue)that can form a covalent bond with the inhibitor.

Further described herein are irreversible inhibitors of other tyrosinekinases, wherein the other tyrosine kinases share homology with Btk.Generally, an irreversible inhibitor compound of Btk used in the methodsdescribed herein is identified or characterized in an in vitro assay,e.g., an acellular biochemical assay or a cellular functional assay.Such assays are useful to determine an in vitro IC₅₀ for an irreversibleBtk inhibitor compound.

For example, an acellular kinase assay can be used to determine Btkactivity after incubation of the kinase in the absence or presence of arange of concentrations of a candidate irreversible Btk inhibitorcompound. If the candidate compound is in fact an irreversible Btkinhibitor, Btk kinase activity will not be recovered by repeat washingwith inhibitor-free medium. See, e.g., J. B. Smaill, et al. (1999), J.Med. Chem. 42(10)):1803-1815. Further, covalent complex formationbetween Btk and a candidate irreversible Btk inhibitor is a usefulindicator of irreversible inhibition of Btk that can be readilydetermined by a number of methods known in the art (e.g., massspectrometry). For example, some irreversible Btk-inhibitor compoundscan form a covalent bond with Cys 481 of Btk (e.g., via a Michaelreaction).

Cellular functional assays for Btk inhibition include measuring one ormore cellular endpoints in response to stimulating a Btk-mediatedpathway in a cell line (e.g., BCR activation in Ramos cells) in theabsence or presence of a range of concentrations of a candidateirreversible Btk inhibitor compound. Useful endpoints for determining aresponse to BCR activation include, e.g., autophosphorylation of Btk,phosphorylation of a Btk target protein (e.g., PLC-γ), and cytoplasmiccalcium flux.

High throughput assays for many acellular biochemical assays (e.g.,kinase assays) and cellular functional assays (e.g., calcium flux) arewell known to those of ordinary skill in the art. In addition, highthroughput screening systems are commercially available (see, e.g.,Zymark Corp., Hopkinton, Mass.; Air Technical Industries, Mentor, Ohio;Beckman Instruments, Inc. Fullerton, Calif.; Precision Systems, Inc.,Natick, Mass., etc.). These systems typically automate entire proceduresincluding all sample and reagent pipetting, liquid dispensing, timedincubations, and final readings of the microplate in detector(s)appropriate for the assay. Automated systems thereby allow theidentification and characterization of a large number of irreversibleBtk compounds without undue effort.

Irreversible Btk inhibitor compounds can be used for the manufacture ofa medicament for treating any of the foregoing conditions (e.g.,autoimmune diseases, inflammatory diseases, allergy disorders, B-cellproliferative disorders, or thromboembolic disorders).

In some embodiments, the irreversible Btk inhibitor compound used forthe methods described herein inhibits Btk or a Btk homolog kinaseactivity with an in vitro IC₅₀ of less than about 10 μM (e.g., less thanabout 1 μM, less than about 0.5 μM, less than about 0.4 μM, less thanabout 0.3 μM, less than about 0.1 μM, less than about 0.08 μM, less thanabout 0.06 μM, less than about 0.05 μM, less than about 0.04 μM, lessthan about 0.03 μM, less than about 0.02 μM, less than about 0.01 μM,less than about 0.008 μM, less than about 0.006 μM, less than about0.005 μM, less than about 0.004 μM, less than about 0.003 μM, less thanabout 0.002 μM, less than about 0.001 μM, less than about 0.00099 μM,less than about 0.00098 μM, less than about 0.00097 μM, less than about0.00096 μM, less than about 0.00095 μM, less than about 0.00094 μM, lessthan about 0.00093 μM, less than about 0.00092 μM, or less than about0.00090 μM).

In one embodiment, the irreversible Btk inhibitor compound selectivelyand irreversibly inhibits an activated form of its target tyrosinekinase (e.g., a phosphorylated form of the tyrosine kinase). Forexample, activated Btk is transphosphorylated at tyrosine 551. Thus, inthese embodiments the irreversible Btk inhibitor inhibits the targetkinase in cells only once the target kinase is activated by thesignaling events.

Described herein are compounds of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). Also described herein are pharmaceutically acceptablesalts, pharmaceutically acceptable solvates, pharmaceutically activemetabolites, and pharmaceutically acceptable prodrugs of such compounds.Pharmaceutical compositions that include at least one such compound or apharmaceutically acceptable salt, pharmaceutically acceptable solvate,pharmaceutically active metabolite or pharmaceutically acceptableprodrug of such compound, are provided. In some embodiments, whencompounds disclosed herein contain an oxidizable nitrogen atom, thenitrogen atom can be converted to an N-oxide by methods well known inthe art. In certain embodiments, isomers and chemically protected formsof compounds having a structure represented by any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) are also provided.

In yet another embodiment, provided herein are kinase inhibitorsaccording to compounds of formula (I).

In another aspect, the present invention provides a compound accordingto Formula (I) having the structure:

wherein:

-   A is

-   Hy is 2-pyridyl substituted with 1-5 groups independently selected    from R⁴, or-   Hy is imidazolyl, thiazolyl, oxazolyl, pyrazolyl, oxadiazolyl,    thiadiazolyl, triazolyl, isothiazolyl, or tetrazolyl, each of which    substituted with 1-2 groups independently selected from R⁴;-   each R¹ and R² is independently H, alkyl, or CN; or R¹ and R²    together form a bond;-   R³ is independently H, alkyl, CN, or cycloalkyl;-   each R⁴ is independently H, halo, hydroxyl, CN, substituted or    unsubstituted alkyl, substituted or unsubstituted amino, substituted    or unsubstituted alkoxy, substituted or unsubstituted amido,    substituted or unsubstituted sulfonyl, substituted or unsubstituted    carboxy, substituted or unsubstituted aryl, or substituted or    unsubstituted heteroaryl; or two adjacent R⁴s connect together with    Hy to form a bicyclic ring optionally substituted with alkyl,    haloalkyl or CN;-   each n is independently 0, 1, or 2; and-   p is 0, 1 or 2;-   or a metabolite, a solvate, a pharmaceutically acceptable salt, or a    prodrug thereof.

In one embodiment, n is 0. In another embodiment, n is 1. In anotherembodiment, n is 2. In one embodiment, p is 0. In another embodiment, pis 1. In another embodiment, p is 2. In one embodiment, n is 0 and p is0. In another embodiment, n is 1 and p is 0. In another embodiment, n is2 and p is 0. In one embodiment, n is 0 and p is 1. In anotherembodiment, n is 1 and p is 1. In another embodiment, n is 2 and p is 1.In one embodiment, n is 0 and p is 2. In another embodiment, n is 1 andp is 2. In another embodiment, n is 2 and p is 2.

In a further embodiment provided herein are compounds according toformula (I):

wherein:

-   Hy is 2-pyridyl substituted with 1-5 groups independently selected    from R⁴, or-   Hy is imidazolyl, thiazolyl, oxazolyl, pyrazolyl, oxadiazolyl,    thiadiazolyl, triazolyl, each of which substituted with 1-2 groups    independently selected from R⁴;-   each R¹ and R² is independently H, alkyl, or CN; or R¹ and R²    together form a bond;-   R³ is independently H, alkyl, CN, or cycloalkyl;-   each R⁴ is independently H, halo, hydroxyl, CN, substituted or    unsubstituted alkyl, substituted or unsubstituted amino, substituted    or unsubstituted alkoxy, substituted or unsubstituted amido,    substituted or unsubstituted sulfonyl, substituted or unsubstituted    carboxy, substituted or unsubstituted aryl, or substituted or    unsubstituted heteroaryl;-   and each n is independently 0, 1, or 2;-   or a metabolite, a solvate, a pharmaceutically acceptable salt, or a    prodrug thereof.

In another embodiment, the compound is according to formula (Ia), (Ib)or (Ic):

wherein Hy, R¹, R², R³, R⁴, and n are as described for formula (I), or ametabolite, a solvate, a pharmaceutically acceptable salt, or a prodrugthereof.

In some embodiments, Hy is 2-pyridyl substituted with 1-5 groupsindependently selected from R⁴, or Hy is imidazolyl, thiazolyl,oxazolyl, pyrazolyl, oxadiazolyl, thiadiazolyl, triazolyl, each of whichsubstituted with 1-2 groups independently selected from R⁴, each R⁴ isindependently H, halo, hydroxyl, CN, substituted or unsubstituted alkyl,substituted or unsubstituted amino, substituted or unsubstituted alkoxy,substituted or unsubstituted amido, substituted or unsubstitutedsulfonyl, substituted or unsubstituted carboxy, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl.

In one embodiment, Hy is 2-pyridyl substituted with 1-5 groupsindependently selected from R⁴.

In another embodiment, Hy is 2-thiazolyl substituted with 1-2 groupsindependently selected from R⁴.

In some embodiments, Hy is selected from:

and R⁴ is as described herein.

In some embodiments, Hy is selected from:

In another embodiment, the compound is according to formula (IIa),(IIb), (IIc), or (IId):

wherein R¹, R², R³, R⁴, and n are as described for formula (I); and m is1, 2, or 3; ora metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In one embodiment, R¹ is H, Me, Et, n-Pr, i-Pr, or CN. In a particularembodiment, R¹ is H, or Me.

In one embodiment, R² is H, Me, Et, n-Pr, or i-Pr.

In another particular embodiment, each R¹ and R² is H.

In a yet another particular embodiment, R¹ and R² together form a bond.

In another embodiment, the compound is according to formula (IIIa),(IIIb), (IIIc) or (IIId):

wherein R³, R⁴, and n are as described for formula (I); ora metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (IIIe),(IIIf), (IIIg) or (IIIh):

wherein R³, R⁴, and n are as described for formula (I); ora metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, n is 0.

In another embodiment, n is 1.

In another embodiment, the compound is according to formula (IVa),(IVb), (IVc), or (IVd):

wherein R³, and R⁴ are as described for formula (I); ora metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (Va), (Vb),(Vc), or (Vd):

wherein R³, and R⁴ are as described for formula (I); ora metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In one embodiment, R³ is H.

In another embodiment, R³ is C₁-C₄ alkyl.

In another embodiment, R³ is Me, Et, n-Pr, or i-Pr.

In another embodiment, R³ is alkoxyalkyl.

In another embodiment, R³ is alkoxymethyl.

In another embodiment, R³ is methoxymethyl

In another embodiment, R³ is hydroxyalkyl.

In another embodiment, R³ is hydroxymethyl.

In another embodiment, R³ is cycloalkyl.

In another embodiment, R³ is cyclopropyl.

In another embodiment, R³ is CN.

In another embodiment, the compound is according to formula (VIa),(VIb), (VIc), or (VId):

wherein R⁴ is as described for formula (I); ora metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (VIe),(VIf), (VIg) or (VIh):

wherein R⁴ is as described for formula (I); ora metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (VIIa),(VIIb), (VIIc) or (VIId):

wherein R⁴ is as described for formula (I); ora metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In one embodiment, R⁴ is H.

In another embodiment, R⁴ is Me, Et, n-Pr, or i-Pr.

In another embodiment, R⁴ is CF₃.

In another embodiment, R⁴ is CN.

In another embodiment, R⁴ is Me.

In another embodiment, R⁴ is cyclopropyl.

In another embodiment, two adjacent R⁴s connect together with Hy to forma bicyclic ring optionally substituted with alkyl, haloalkyl or CN.

In another embodiment, the compound is according to formula (VIIIa),(VIIIb), (VIIIc), or (VIIId):

or a metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (VIIIe),(VIIIf), (VIIIg) or (VIIIh):

or a metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (IXa),(IXb), (IXc), or (IXd):

or a metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (Xa), (Xb),(Xc), or (Xd):

or a metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (Xe), (Xf),(Xg), or (Xh):

or a metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (XIa),(XIb), (XIc), or (XId):

or a metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (XIe),(XIf), (XIg), (XIh), (XIi) or (XIj):

or a metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is according to formula (XIIa),(XIIb), (XIIc), (XIId), (XIIe), (XIIf), (XIIg), or (XIIh):

or a metabolite, a solvate, a pharmaceutically acceptable salt, or aprodrug thereof.

In another embodiment, the compound is any one of the compounds selectedfrom the compounds listed in Table 1.

In one embodiment, the compound is compound VIIIa.

In another embodiment, the compound is compound VIIIb.

In another embodiment, the compound is compound VIIIc.

In another embodiment, the compound is compound VIIId.

In one embodiment, the compound is compound VIIIe.

In another embodiment, the compound is compound VIIIf.

In another embodiment, the compound is compound VIIIg.

In another embodiment, the compound is compound VIIIh.

In one embodiment, the compound is compound IXa.

In another embodiment, the compound is compound IXb.

In another embodiment, the compound is compound IXc.

In another embodiment, the compound is compound IXd.

In one embodiment, the compound is compound Xa.

In another embodiment, the compound is compound Xb.

In another embodiment, the compound is compound Xc.

In another embodiment, the compound is compound Xd.

In one embodiment, the compound is compound Xe.

In another embodiment, the compound is compound Xf.

In another embodiment, the compound is compound Xg.

In another embodiment, the compound is compound Xh.

In one embodiment, the compound is compound XIa.

In another embodiment, the compound is compound XIb.

In another embodiment, the compound is compound XIc.

In another embodiment, the compound is compound XId.

In one embodiment, the compound is compound XIe.

In another embodiment, the compound is compound XIf.

In another embodiment, the compound is compound XIg.

In another embodiment, the compound is compound XIh.

In one embodiment, the compound is compound XIi.

In another embodiment, the compound is compound XIj.

Embodiments of the compounds of Formula (I) displayed improved potencyagainst Btk with IC₅₀ values of as low as less than 1 nM or less than0.1 nM, and/or high occupancy of active site of Btk (e.g., more than50%, 70% or 90% occupancy) at low dosages of below 5 mg/kg (e.g., at orbelow 3 mg/kg) when administered in vivo (e.g., in rats).

In another aspect, the present invention provides, a pharmaceuticalcomposition comprising a compound according to formula (I).

In yet another aspect, the present invention provides, a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundof formula (I), and a pharmaceutically acceptable excipient.

In one embodiment, the pharmaceutical composition is formulated for aroute of administration selected from oral administration, parenteraladministration, buccal administration, nasal administration, topicaladministration, or rectal administration.

In yet another aspect, the present invention provides, methods fortreating an autoimmune disease or condition comprising administering toa patient in need the pharmaceutical composition of the presentinvention.

In one embodiment, the autoimmune disease is selected from rheumatoidarthritis or lupus.

In yet another aspect, the present invention provides, methods fortreating a heteroimmune disease or condition comprising administering toa patient in need the pharmaceutical composition of the presentinvention.

In yet another aspect, the present invention provides, for treating acancer comprising administering to a patient in need the pharmaceuticalcomposition the present invention.

In one embodiment, the cancer is a B-cell proliferative disorder.

In another embodiment, the B-cell proliferative disorder is diffuselarge B cell lymphoma, follicular lymphoma or chronic lymphocyticleukemia.

In yet another aspect, the present invention provides, methods fortreating mastocytosis comprising administering to a patient in need thepharmaceutical composition of the present invention.

In yet another aspect, the present invention provides, methods fortreating osteoporosis or bone resorption disorders comprisingadministering to a patient in need the pharmaceutical composition of thepresent invention.

In yet another aspect, the present invention provides, methods ortreating an inflammatory disease or condition comprising administeringto a patient in need the pharmaceutical composition of the presentinvention.

In yet another aspect, the present invention provides, methods fortreating lupus comprising administering to a subject in need thereof acomposition containing a therapeutically effective amount of a compoundof formula (I) that is inhibitor of the Bruton's tyrosine kinase or aBruton's tyrosine kinase homolog.

In yet another aspect, the present invention provides, methods fortreating a heteroimmune disease or condition comprising administering toa subject in need thereof a composition containing a therapeuticallyeffective amount of a compound of formula (I) that is inhibitor of theBruton's tyrosine kinase or a Bruton's tyrosine kinase homolog.

In yet another aspect, the present invention provides, methods fortreating diffuse large B cell lymphoma, follicular lymphoma or chroniclymphocytic leukemia comprising administering to a subject in needthereof a composition containing a therapeutically effective amount of acompound of formula (I) that is inhibitor of the Bruton's tyrosinekinase or a Bruton's tyrosine kinase homolog.

In yet another aspect, the present invention provides, methods fortreating mastocytosis, comprising administering to a subject in needthereof a composition containing a therapeutically effective amount of acompound of formula (I) that is inhibitor of the Bruton's tyrosinekinase or a Bruton's tyrosine kinase homolog.

In yet another aspect, the present invention provides, methods fortreating osteoporosis or bone resorption disorders comprisingadministering to a subject in need thereof a composition containing atherapeutically effective amount of a compound of formula (I) that isinhibitor of the Bruton's tyrosine kinase or a Bruton's tyrosine kinasehomolog.

In yet another aspect, the present invention provides, methods fortreating an inflammatory disease or condition comprising administeringto a subject in need thereof a composition containing a therapeuticallyeffective amount of a compound of formula (I) that is inhibitor of theBruton's tyrosine kinase or a Bruton's tyrosine kinase homolog.

In one embodiment, the kinase inhibitor binds to at least one of Btk,Jak3, Blk, Bmx, Tec, and Itk.

In a particular embodiment, the kinase inhibitor binds to Btk.

In another particular embodiment, the kinase inhibitor binds to Jak3.

In another particular embodiment, the kinase inhibitor binds to Tec.

In another particular embodiment, the kinase inhibitor binds to Btk andTec.

In another particular embodiment, the kinase inhibitor binds to Blk.

In another particular embodiment, the kinase inhibitor binds to amultiplicity of src-family protein kinase inhibitors.

In another particular embodiment, the compound is:

-   (R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-pent-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-(3-cyclopropylpropioloyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-(4-methoxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-(4-hydroxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide;-   R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide;-   (R,E)-4-(4-amino-1-(1-but-2-enoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-(3-methylbut-2-enoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide;-   4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;-   4-(1-((1R,4R)-4-acrylamidocyclohexyl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;-   4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide;-   4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide;-   4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide;-   4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide;-   4-(1-((1R,4R)-4-acrylamidocyclohexyl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide;-   (R,E)-4-(4-amino-1-(1-pent-2-enoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-methacryloylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R,Z)-4-(4-amino-1-(1-(2-methylbut-2-enoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-(2-cyano-3-methylbut-2-enoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-(4-hydroxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;-   (R)-4-(4-amino-1-(1-(4-methoxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;-   (R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(3-methylisothiazol-5-yl)benzamide;    or-   (R)-4-(4-amino-1-(1-(1-cyanocyclopropanecarbonyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;    or a solvate or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides, a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and apharmaceutically effective amount of a compound according to any one ofthe formulas described herein. In one embodiment, the compound isaccording to any one of formulas: (I), (Ia)-(Ic), (IIa)-(IId),(IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId),(VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh).

In one embodiment, the pharmaceutical composition is formulated for aroute of administration selected from oral administration, parenteraladministration, buccal administration, nasal administration, topicaladministration, or rectal administration.

In one embodiment, the carrier is a parenteral carrier.

In another embodiment, the carrier is an oral carrier.

In another embodiment, the carrier is a topical carrier.

Any combination of the groups described above for the various variablesis contemplated herein. It is understood that substituents andsubstitution patterns on the compounds provided herein can be selectedby one of ordinary skill in the art to provide compounds that arechemically stable and that can be synthesized by techniques known in theart, as well as those set forth herein.

Further embodiments of compounds of Formula (I), (Ia)-(Ic), (IIa)-(IId),(IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId),(VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh)include, but are not limited to, compounds selected from the groupconsisting of compounds with IDs 1-48.

Throughout the specification, groups and substituents thereof can bechosen by one skilled in the field to provide stable moieties andcompounds.

In some embodiments, the compounds of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) reversibly inhibit Btk and in other embodiments are usedto treat patients suffering from Bruton's tyrosine kinase-dependent orBruton's tyrosine kinase mediated conditions or diseases, including, butnot limited to, cancer, autoimmune and other inflammatory diseases.

In some embodiments, the compounds of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) inhibit Btk and in other embodiments are used to treatpatients suffering from Bruton's tyrosine kinase-dependent or Bruton'styrosine kinase mediated conditions or diseases, including, but notlimited to, cancer, autoimmune and other inflammatory diseases.

Preparation of Compounds

Compounds of any of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) may be synthesizedusing standard synthetic reactions known to those of skill in the art orusing methods known in the art. The reactions can be employed in alinear sequence to provide the compounds or they may be used tosynthesize fragments which are subsequently joined by the methods knownin the art.

Described herein are compounds that inhibit the activity of tyrosinekinase(s), such as Btk, and processes for their preparation. Alsodescribed herein are pharmaceutically acceptable salts, pharmaceuticallyacceptable solvates, pharmaceutically active metabolites andpharmaceutically acceptable prodrugs of such compounds. Pharmaceuticalcompositions that include at least one such compound or apharmaceutically acceptable salt, pharmaceutically acceptable solvate,pharmaceutically active metabolite or pharmaceutically acceptableprodrug of such compound, are provided.

The starting material used for the synthesis of the compounds describedherein may be synthesized or can be obtained from commercial sources,such as, but not limited to, Aldrich Chemical Co. (Milwaukee, Wis.),Bachem (Torrance, Calif.), or Sigma Chemical Co. (St. Louis, Mo.). Thecompounds described herein, and other related compounds having differentsubstituents can be synthesized using techniques and materials known tothose of skill in the art, such as described, for example, in March,ADVANCED ORGANIC CHEMISTRY 4^(th) Ed., (Wiley 1992); Carey and Sundberg,ADVANCED ORGANIC CHEMISTRY 4^(th) Ed., Vols. A and B (Plenum 2000,2001); Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3^(rd)Ed., (Wiley 1999); Fieser and Fieser's Reagents for Organic Synthesis,Volumes 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of CarbonCompounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers,1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991); andLarock's Comprehensive Organic Transformations (VCH Publishers Inc.,1989). (all of which are incorporated by reference in their entirety).Other methods for the synthesis of compounds described herein may befound in International Patent Publication No. WO 01/01982901, Arnold etal. Bioorganic & Medicinal Chemistry Letters 10 (2000) 2167-2170;Burchat et al. Bioorganic & Medicinal Chemistry Letters 12 (2002)1687-1690. General methods for the preparation of compound as disclosedherein may be derived from known reactions in the field, and thereactions may be modified by the use of appropriate reagents andconditions, as would be recognized by the skilled person, for theintroduction of the various moieties found in the formulae as providedherein.

The products of the reactions may be isolated and purified, if desired,using conventional techniques, including, but not limited to,filtration, distillation, crystallization, chromatography and the like.Such materials may be characterized using conventional means, includingphysical constants and spectral data.

Compounds described herein may be prepared as a single isomer or amixture of isomers.

In some embodiments, compounds of Formula (I) are prepared according togeneral Scheme I. In Scheme I, 4-bromobenzoic acid (A-1) is coupled withamine Hy-NH₂ under conditions generally known in the art (e.g., in thepresence of an amino acid coupling reagent, such as POCl₃), wherein Hyis as defined herein, followed by reaction with a boron reagent, such as[B(OR¹¹)₂]₂ under catalytic conditions such as in the presence of apalladium catalyst, to give Compound A-2, wherein each R¹¹ isindependently H, alkyl or the two R¹¹ form an optionally substitutedalkylene group. Compound A-3 reacts with HO-A-PG such as under Mitsunobucoupling conditions to provide for Compound A-4, wherein A is as definedherein, LG is a leaving group, such as Br or I, and PG is an aminoprotecting group, such as t-butyloxycarbonyl (Boc), mesitylenesulfonyl(Mts), benzyloxycarbonyl (Cbz or Z), 2-chlorobenzyloxycarbonyl,t-butyldimethylsilyl (TBS or TBDMS), 9-fluorenylmethyloxycarbonyl(Fmoc), attached to the nitrogen atom of A. Compound A-4 is deprotectedto provide for Compound A-5. Many amino protecting groups and respectivemethods of deprotection are also known in the art. Compound A-6 is thencoupled with Compound A-7, wherein R¹² is OH or halo, or C(O)R¹²together is an active ester or anhydride, under conditions generallyknown in the art (e.g., in the presence of an amino acid couplingreagent when R¹² is OH), to give a compound of Formula (I). Many activeesters and anhydride groups are known in the art.

Further Forms of Compounds

Compounds disclosed herein have a structure of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). It is understood that when reference is made to compoundsdescribed herein, it is meant to include compounds of any of Formula(I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) as well as to all of the specific compoundsthat fall within the scope of these generic formulae, unless otherwiseindicated.

The compounds described herein may possess one or more stereocenters andeach center may exist in the R or S configuration. The compoundspresented herein include all diastereomeric, enantiomeric, and epimericforms as well as the appropriate mixtures thereof. Stereoisomers may beobtained, if desired, by methods known in the art as, for example, theseparation of stereoisomers by chiral chromatographic columns.

Diastereomeric mixtures can be separated into their individualdiastereomers on the basis of their physical chemical differences bymethods known, for example, by chromatography and/or fractionalcrystallization. In one embodiment, enantiomers can be separated bychiral chromatographic columns. In other embodiments, enantiomers can beseparated by converting the enantiomeric mixture into a diastereomericmixture by reaction with an appropriate optically active compound (e.g.,alcohol), separating the diastereomers and converting (e.g.,hydrolyzing) the individual diastereomers to the corresponding pureenantiomers. All such isomers, including diastereomers, enantiomers, andmixtures thereof are considered as part of the compositions describedherein.

The methods and formulations described herein include the use ofN-oxides, crystalline forms (also known as polymorphs), orpharmaceutically acceptable salts of compounds described herein, as wellas active metabolites of these compounds having the same type ofactivity. In some situations, compounds may exist as tautomers. Alltautomers are included within the scope of the compounds presentedherein. In addition, the compounds described herein can exist inunsolvated as well as solvated forms with pharmaceutically acceptablesolvents such as water, ethanol, and the like. The solvated forms of thecompounds presented herein are also considered to be disclosed herein.

Compounds of any of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) in unoxidized formcan be prepared from N-oxides of compounds of any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) by treating with a reducing agent, such as,but not limited to, sulfur, sulfur dioxide, triphenyl phosphine, lithiumborohydride, sodium borohydride, phosphorus trichloride, tribromide, orthe like in a suitable inert organic solvent, such as, but not limitedto, acetonitrile, ethanol, aqueous dioxane, or the like at 0 to 80° C.

In some embodiments, compounds described herein are prepared asprodrugs. A “prodrug” refers to an agent that is converted into theparent drug in vivo. Prodrugs are often useful because, in somesituations, they may be easier to administer than the parent drug. Theymay, for instance, be bioavailable by oral administration whereas theparent is not. The prodrug may also have improved solubility inpharmaceutical compositions over the parent drug. An example, withoutlimitation, of a prodrug would be a compound described herein, which isadministered as an ester (the “prodrug”) to facilitate transmittalacross a cell membrane where water solubility is detrimental to mobilitybut which then is metabolically hydrolyzed to the carboxylic acid, theactive entity, once inside the cell where water-solubility isbeneficial. A further example of a prodrug might be a short peptide(polyaminoacid) bonded to an acid group where the peptide is metabolizedto reveal the active moiety. In certain embodiments, upon in vivoadministration, a prodrug is chemically converted to the biologically,pharmaceutically or therapeutically active form of the compound. Incertain embodiments, a prodrug is enzymatically metabolized by one ormore steps or processes to the biologically, pharmaceutically ortherapeutically active form of the compound. To produce a prodrug, apharmaceutically active compound is modified such that the activecompound will be regenerated upon in vivo administration. The prodrugcan be designed to alter the metabolic stability or the transportcharacteristics of a drug, to mask side effects or toxicity, to improvethe flavor of a drug or to alter other characteristics or properties ofa drug. By virtue of knowledge of pharmacodynamic processes and drugmetabolism in vivo, those of skill in this art, once a pharmaceuticallyactive compound is known, can design prodrugs of the compound. (see, forexample, Nogrady (1985) Medicinal Chemistry A Biochemical Approach,Oxford University Press, New York, pages 388-392; Silverman (1992), TheOrganic Chemistry of Drug Design and Drug Action, Academic Press, Inc.,San Diego, pages 352-401, Saulnier et al., (1994), Bioorganic andMedicinal Chemistry Letters, Vol. 4, p. 1985).

Prodrug forms of the herein described compounds, wherein the prodrug ismetabolized in vivo to produce a derivative as set forth herein areincluded within the scope of the claims. In some cases, some of theherein-described compounds may be a prodrug for another derivative oractive compound.

Prodrugs are often useful because, in some situations, they may beeasier to administer than the parent drug. They may, for instance, bebioavailable by oral administration whereas the parent is not. Theprodrug may also have improved solubility in pharmaceutical compositionsover the parent drug. Prodrugs may be designed as reversible drugderivatives, for use as modifiers to enhance drug transport tosite-specific tissues. In some embodiments, the design of a prodrugincreases the effective water solubility. See, e.g., Fedorak et al., Am.J. Physiol., 269:G210-218 (1995); McLoed et al., Gastroenterol,106:405-413 (1994); Hochhaus et al., Biomed. Chrom., 6:283-286 (1992);J. Larsen and H. Bundgaard, Int. J. Pharmaceutics, 37, 87 (1987); J.Larsen et al., Int. J. Pharmaceutics, 47, 103 (1988); Sinkula et al., J.Pharm. Sci., 64:181-210 (1975); T. Higuchi and V. Stella, Pro-drugs asNovel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series; andEdward B. Roche, Bioreversible Carriers in Drug Design, AmericanPharmaceutical Association and Pergamon Press, 1987, all incorporatedherein in their entirety.

Sites on the aromatic ring portion of compounds of any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) can be susceptible to various metabolicreactions, therefore incorporation of appropriate substituents on thearomatic ring structures, such as, by way of example only, halogens canreduce, minimize or eliminate this metabolic pathway.

Compounds described herein include isotopically-labeled compounds, whichare identical to those recited in the various formulas and structurespresented herein, but for the fact that one or more atoms are replacedby an atom having an atomic mass or mass number different from theatomic mass or mass number usually found in nature. Examples of isotopesthat can be incorporated into the present compounds include isotopes ofhydrogen, carbon, nitrogen, oxygen, fluorine and chlorine, such as ²H,³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³⁵S, ¹⁸F, ³⁶Cl, respectively. Certainisotopically-labeled compounds described herein, for example those intowhich radioactive isotopes such as ³H and ¹⁴C are incorporated, areuseful in drug and/or substrate tissue distribution assays. Further,substitution with isotopes such as deuterium, i.e., ²H, can affordcertain therapeutic advantages resulting from greater metabolicstability, for example increased in vivo half-life or reduced dosagerequirements.

In additional or further embodiments, the compounds described herein aremetabolized upon administration to an organism in need to produce ametabolite that is then used to produce a desired effect, including adesired therapeutic effect.

Compounds described herein may be formed as, and/or used as,pharmaceutically acceptable salts. The type of pharmaceutical acceptablesalts, include, but are not limited to: (1) acid addition salts, formedby reacting the free base form of the compound with a pharmaceuticallyacceptable: inorganic acid such as hydrochloric acid, hydrobromic acid,sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, andthe like; or with an organic acid such as acetic acid, propionic acid,hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid,lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, trifluoroacetic acid, tartaric acid, citric acid, benzoicacid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonicacid, 2-naphthalenesulfonic acid,4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid,4,4′-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionicacid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuricacid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylicacid, stearic acid, muconic acid, and the like; (2) salts formed when anacidic proton present in the parent compound either is replaced by ametal ion, e.g., an alkali metal ion (e.g. lithium, sodium, potassium),an alkaline earth ion (e.g. magnesium, or calcium), or an aluminum ion;or coordinates with an organic base. Acceptable organic bases includeethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like. Acceptable inorganic bases includealuminum hydroxide, calcium hydroxide, potassium hydroxide, sodiumcarbonate, sodium hydroxide, and the like.

The corresponding counterions of the pharmaceutically acceptable saltsmay be analyzed and identified using various methods including, but notlimited to, ion exchange chromatography, ion chromatography, capillaryelectrophoresis, inductively coupled plasma, atomic absorptionspectroscopy, mass spectrometry, or any combination thereof.

The salts are recovered by using at least one of the followingtechniques: filtration, precipitation with a non-solvent followed byfiltration, evaporation of the solvent, or, in the case of aqueoussolutions, lyophilization.

It should be understood that a reference to a pharmaceuticallyacceptable salt includes the solvent addition forms or crystal formsthereof, particularly solvates or polymorphs. Solvates contain eitherstoichiometric or non-stoichiometric amounts of a solvent, and may beformed during the process of crystallization with pharmaceuticallyacceptable solvents such as water, ethanol, and the like. Hydrates areformed when the solvent is water, or alcoholates are formed when thesolvent is alcohol. Solvates of compounds described herein can beconveniently prepared or formed during the processes described herein.In addition, the compounds provided herein can exist in unsolvated aswell as solvated forms. In general, the solvated forms are consideredequivalent to the unsolvated forms for the purposes of the compounds andmethods provided herein.

It should be understood that a reference to a salt includes the solventaddition forms or crystal forms thereof, particularly solvates orpolymorphs. Solvates contain either stoichiometric or non-stoichiometricamounts of a solvent, and are often formed during the process ofcrystallization with pharmaceutically acceptable solvents such as water,ethanol, and the like. Hydrates are formed when the solvent is water, oralcoholates are formed when the solvent is alcohol. Polymorphs includethe different crystal packing arrangements of the same elementalcomposition of a compound. Polymorphs usually have different X-raydiffraction patterns, infrared spectra, melting points, density,hardness, crystal shape, optical and electrical properties, stability,and solubility. Various factors such as the recrystallization solvent,rate of crystallization, and storage temperature may cause a singlecrystal form to dominate.

Compounds described herein may be in various forms, including but notlimited to, amorphous forms, milled forms and nano-particulate forms. Inaddition, compounds described herein include crystalline forms, alsoknown as polymorphs. Polymorphs include the different crystal packingarrangements of the same elemental composition of a compound. Polymorphsusually have different X-ray diffraction patterns, infrared spectra,melting points, density, hardness, crystal shape, optical and electricalproperties, stability, and solubility. Various factors such as therecrystallization solvent, rate of crystallization, and storagetemperature may cause a single crystal form to dominate.

The screening and characterization of the pharmaceutically acceptablesalts, polymorphs and/or solvates may be accomplished using a variety oftechniques including, but not limited to, thermal analysis, x-raydiffraction, spectroscopy, vapor sorption, and microscopy. Thermalanalysis methods address thermo chemical degradation or thermo physicalprocesses including, but not limited to, polymorphic transitions, andsuch methods are used to analyze the relationships between polymorphicforms, determine weight loss, to find the glass transition temperature,or for excipient compatibility studies. Such methods include, but arenot limited to, Differential scanning calorimetry (DSC), ModulatedDifferential Scanning calorimetry (MDCS), Thermogravimetric analysis(TGA), and Thermogravi-metric and Infrared analysis (TG/IR). X-raydiffraction methods include, but are not limited to, single crystal andpowder diffractometers and synchrotron sources. The variousspectroscopic techniques used include, but are not limited to, Raman,FTIR, UVIS, and NMR (liquid and solid state). The various microscopytechniques include, but are not limited to, polarized light microscopy,Scanning Electron Microscopy (SEM) with Energy Dispersive X-Ray Analysis(EDX), Environmental Scanning Electron Microscopy with EDX (in gas orwater vapor atmosphere), IR microscopy, and Raman microscopy.

Throughout the specification, groups and substituents thereof can bechosen by one skilled in the field to provide stable moieties andcompounds.

Pharmaceutical Composition/Formulation

Pharmaceutical compositions may be formulated in a conventional mannerusing one or more physiologically acceptable carriers includingexcipients and auxiliaries which facilitate processing of the activecompounds into preparations which can be used pharmaceutically. Properformulation is dependent upon the route of administration chosen. Any ofthe well-known techniques, carriers, and excipients may be used assuitable and as understood in the art. A summary of pharmaceuticalcompositions described herein may be found, for example, in Remington:The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: MackPublishing Company, 1995); Hoover, John E., Remington's PharmaceuticalSciences, Mack Publishing Co., Easton, Pa. 1975; Liberman, H. A. andLachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York,N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems,Seventh Ed. (Lippincott Williams & Wilkins 1999), herein incorporated byreference in their entirety.

A pharmaceutical composition, as used herein, refers to a mixture of acompound described herein, such as, for example, compounds of any ofFormula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) with other chemical components,such as carriers, stabilizers, diluents, dispersing agents, suspendingagents, thickening agents, and/or excipients. The pharmaceuticalcomposition facilitates administration of the compound to an organism.In practicing the methods of treatment or use provided herein,therapeutically effective amounts of compounds described herein areadministered in a pharmaceutical composition to a mammal having adisease, disorder, or condition to be treated. Preferably, the mammal isa human. A therapeutically effective amount can vary widely depending onthe severity of the disease, the age and relative health of the subject,the potency of the compound used and other factors. The compounds can beused singly or in combination with one or more therapeutic agents ascomponents of mixtures.

In certain embodiments, compositions may also include one or more pHadjusting agents or buffering agents, including acids such as acetic,boric, citric, lactic, phosphoric and hydrochloric acids; bases such assodium hydroxide, sodium phosphate, sodium borate, sodium citrate,sodium acetate, sodium lactate and tris-hydroxymethylaminomethane; andbuffers such as citrate/dextrose, sodium bicarbonate and ammoniumchloride. Such acids, bases and buffers are included in an amountrequired to maintain pH of the composition in an acceptable range.

In other embodiments, compositions may also include one or more salts inan amount required to bring osmolality of the composition into anacceptable range. Such salts include those having sodium, potassium orammonium cations and chloride, citrate, ascorbate, borate, phosphate,bicarbonate, sulfate, thiosulfate or bisulfite anions; suitable saltsinclude sodium chloride, potassium chloride, sodium thiosulfate, sodiumbisulfite and ammonium sulfate.

The term “pharmaceutical combination” as used herein, means a productthat results from the mixing or combining of more than one activeingredient and includes both fixed and non-fixed combinations of theactive ingredients. The term “fixed combination” means that the activeingredients, e.g. a compound described herein and a co-agent, are bothadministered to a patient simultaneously in the form of a single entityor dosage. The term “non-fixed combination” means that the activeingredients, e.g. a compound described herein and a co-agent, areadministered to a patient as separate entities either simultaneously,concurrently or sequentially with no specific intervening time limits,wherein such administration provides effective levels of the twocompounds in the body of the patient. The latter also applies tococktail therapy, e.g. the administration of three or more activeingredients.

The pharmaceutical compositions described herein can be administered toa subject by multiple administration routes, including but not limitedto, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular),intranasal, buccal, topical, rectal, or transdermal administrationroutes. The pharmaceutical compositions described herein include, butare not limited to, aqueous liquid dispersions, self-emulsifyingdispersions, solid solutions, liposomal dispersions, aerosols, soliddosage forms, powders, immediate release formulations, controlledrelease formulations, fast melt formulations, tablets, capsules, pills,delayed release formulations, extended release formulations, pulsatilerelease formulations, multiparticulate formulations, and mixed immediateand controlled release formulations.

Pharmaceutical compositions including a compound described herein may bemanufactured in a conventional manner, such as, by way of example only,by means of conventional mixing, dissolving, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or compressionprocesses.

The pharmaceutical compositions will include at least one compounddescribed herein, such as, for example, a compound of any of Formula(I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) as an active ingredient in free-acid orfree-base form, or in a pharmaceutically acceptable salt form. Inaddition, the methods and pharmaceutical compositions described hereininclude the use of N-oxides, crystalline forms (also known aspolymorphs), as well as active metabolites of these compounds having thesame type of activity. In some situations, compounds may exist astautomers. All tautomers are included within the scope of the compoundspresented herein. Additionally, the compounds described herein can existin unsolvated as well as solvated forms with pharmaceutically acceptablesolvents such as water, ethanol, and the like. The solvated forms of thecompounds presented herein are also considered to be disclosed herein.

“Antifoaming agents” reduce foaming during processing which can resultin coagulation of aqueous dispersions, bubbles in the finished film, orgenerally impair processing. Exemplary anti-foaming agents includesilicon emulsions or sorbitan sesquoleate.

“Antioxidants” include, for example, butylated hydroxytoluene (BHT),sodium ascorbate, ascorbic acid, sodium metabisulfite and tocopherol. Incertain embodiments, antioxidants enhance chemical stability whererequired.

In certain embodiments, compositions provided herein may also includeone or more preservatives to inhibit microbial activity. Suitablepreservatives include mercury-containing substances such as merfen andthiomersal; stabilized chlorine dioxide; and quaternary ammoniumcompounds such as benzalkonium chloride, cetyltrimethylammonium bromideand cetylpyridinium chloride.

Formulations described herein may benefit from antioxidants, metalchelating agents, thiol containing compounds and other generalstabilizing agents. Examples of such stabilizing agents, include, butare not limited to: (a) about 0.5% to about 2% w/v glycerol, (b) about0.1% to about 1% w/v methionine, (c) about 0.1% to about 2% w/vmonothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% toabout 2% w/v ascorbic acid, (f) 0.003% to about 0.02% w/v polysorbate80, (g) 0.001% to about 0.05% w/v. polysorbate 20, (h) arginine, (i)heparin, (j) dextran sulfate, (k) cyclodextrins, (1) pentosanpolysulfate and other heparinoids, (m) divalent cations such asmagnesium and zinc; or (n) combinations thereof.

“Binders” impart cohesive qualities and include, e.g., alginic acid andsalts thereof; cellulose derivatives such as carboxymethylcellulose,methylcellulose (e.g., Methocel®), hydroxypropylmethylcellulose,hydroxyethylcellulose, hydroxypropylcellulose (e.g., Klucel®),ethylcellulose (e.g., Ethocel®), and microcrystalline cellulose (e.g.,Avicel®); microcrystalline dextrose; amylose; magnesium aluminumsilicate; polysaccharide acids; bentonites; gelatin;polyvinylpyrrolidone/vinyl acetate copolymer; crosspovidone; povidone;starch; pregelatinized starch; tragacanth, dextrin, a sugar, such assucrose (e.g., Dipac®), glucose, dextrose, molasses, mannitol, sorbitol,xylitol (e.g., Xylitab®), and lactose; a natural or synthetic gum suchas acacia, tragacanth, ghatti gum, mucilage of isapol husks,polyvinylpyrrolidone (e.g., Polyvidone® CL, Kollidon® CL, Polyplasdone®XL-10), larch arabogalactan, Veegum®, polyethylene glycol, waxes, sodiumalginate, and the like.

A “carrier” or “carrier materials” include any commonly used excipientsin pharmaceutics and should be selected on the basis of compatibilitywith compounds disclosed herein, such as, compounds of any of Formula(I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) and the release profile properties of thedesired dosage form. Exemplary carrier materials include, e.g., binders,suspending agents, disintegration agents, filling agents, surfactants,solubilizers, stabilizers, lubricants, wetting agents, diluents, and thelike. “Pharmaceutically compatible carrier materials” may include, butare not limited to, acacia, gelatin, colloidal silicon dioxide, calciumglycerophosphate, calcium lactate, maltodextrin, glycerine, magnesiumsilicate, polyvinylpyrrollidone (PVP), cholesterol, cholesterol esters,sodium caseinate, soy lecithin, taurocholic acid, phosphotidylcholine,sodium chloride, tricalcium phosphate, dipotassium phosphate, celluloseand cellulose conjugates, sugars sodium stearoyl lactylate, carrageenan,monoglyceride, diglyceride, pregelatinized starch, and the like. See,e.g., Remington: The Science and Practice of Pharmacy, Nineteenth Ed(Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E.,Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical DosageForms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams &Wilkins 1999).

“Dispersing agents,” and/or “viscosity modulating agents” includematerials that control the diffusion and homogeneity of a drug throughliquid media or a granulation method or blend method. In someembodiments, these agents also facilitate the effectiveness of a coatingor eroding matrix. Exemplary diffusion facilitators/dispersing agentsinclude, e.g., hydrophilic polymers, electrolytes, Tween® 60 or 80, PEG,polyvinylpyrrolidone (PVP; commercially known as Plasdone®), and thecarbohydrate-based dispersing agents such as, for example, hydroxypropylcelluloses (e.g., HPC, HPC-SL, and HPC-L), hydroxypropylmethylcelluloses (e.g., HPMC K100, HPMC K4M, HPMC K15M, and HPMC K100M),carboxymethylcellulose sodium, methylcellulose, hydroxyethylcellulose,hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate,hydroxypropylmethylcellulose acetate stearate (HPMCAS), noncrystallinecellulose, magnesium aluminum silicate, triethanolamine, polyvinylalcohol (PVA), vinyl pyrrolidone/vinyl acetate copolymer (S630),4-(1,1,3,3-tetramethylbutyl)-phenol polymer with ethylene oxide andformaldehyde (also known as tyloxapol), poloxamers (e.g., PluronicsF68®, F88®, and F108®, which are block copolymers of ethylene oxide andpropylene oxide); and poloxamines (e.g., Tetronic 908®, also known asPoloxamine 908®, which is a tetrafunctional block copolymer derived fromsequential addition of propylene oxide and ethylene oxide toethylenediamine (BASF Corporation, Parsippany, N.J.)),polyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidoneK25, or polyvinylpyrrolidone K30, polyvinylpyrrolidone/vinyl acetatecopolymer (S-630), polyethylene glycol, e.g., the polyethylene glycolcan have a molecular weight of about 300 to about 6000, or about 3350 toabout 4000, or about 7000 to about 5400, sodium carboxymethylcellulose,methylcellulose, polysorbate-80, sodium alginate, gums, such as, e.g.,gum tragacanth and gum acacia, guar gum, xanthans, including xanthangum, sugars, cellulosics, such as, e.g., sodium carboxymethylcellulose,methylcellulose, sodium carboxymethylcellulose, polysorbate-80, sodiumalginate, polyethoxylated sorbitan monolaurate, polyethoxylated sorbitanmonolaurate, povidone, carbomers, polyvinyl alcohol (PVA), alginates,chitosans and combinations thereof. Plasticizers such as cellulose ortriethyl cellulose can also be used as dispersing agents. Dispersingagents particularly useful in liposomal dispersions and self-emulsifyingdispersions are dimyristoyl phosphatidyl choline, natural phosphatidylcholine from eggs, natural phosphatidyl glycerol from eggs, cholesteroland isopropyl myristate.

Combinations of one or more erosion facilitator with one or morediffusion facilitator can also be used in the present compositions.

The term “diluent” refers to chemical compounds that are used to dilutethe compound of interest prior to delivery. Diluents can also be used tostabilize compounds because they can provide a more stable environment.Salts dissolved in buffered solutions (which also can provide pH controlor maintenance) are utilized as diluents in the art, including, but notlimited to a phosphate buffered saline solution. In certain embodiments,diluents increase bulk of the composition to facilitate compression orcreate sufficient bulk for homogenous blend for capsule filling. Suchcompounds include e.g., lactose, starch, mannitol, sorbitol, dextrose,microcrystalline cellulose such as Avicel®; dibasic calcium phosphate,dicalcium phosphate dihydrate; tricalcium phosphate, calcium phosphate;anhydrous lactose, spray-dried lactose; pregelatinized starch,compressible sugar, such as Di-Pac® (Amstar); mannitol,hydroxypropylmethylcellulose, hydroxypropylmethylcellulose acetatestearate, sucrose-based diluents, confectioner's sugar; monobasiccalcium sulfate monohydrate, calcium sulfate dihydrate; calcium lactatetrihydrate, dextrates; hydrolyzed cereal solids, amylose; powderedcellulose, calcium carbonate; glycine, kaolin; mannitol, sodiumchloride; inositol, bentonite, and the like.

The term “disintegrate” includes both the dissolution and dispersion ofthe dosage form when contacted with gastrointestinal fluid.“Disintegration agents or disintegrants” facilitate the breakup ordisintegration of a substance. Examples of disintegration agents includea starch, e.g., a natural starch such as corn starch or potato starch, apregelatinized starch such as National 1551 or Amijel®, or sodium starchglycolate such as Promogel® or Explotab®, a cellulose such as a woodproduct, methylcrystalline cellulose, e.g., Avicel®, Avicel® PH101,Avicel® PH102, Avicel® PH105, Elcema® P100, Emcocel®, Vivacel®, MingTia®, and Solka-Floc®, methylcellulose, croscarmellose, or across-linked cellulose, such as cross-linked sodiumcarboxymethylcellulose (Ac-Di-Sol®), cross-linkedcarboxymethylcellulose, or cross-linked croscarmellose, a cross-linkedstarch such as sodium starch glycolate, a cross-linked polymer such ascrosspovidone, a cross-linked polyvinylpyrrolidone, alginate such asalginic acid or a salt of alginic acid such as sodium alginate, a claysuch as Veegum® HV (magnesium aluminum silicate), a gum such as agar,guar, locust bean, Karaya, pectin, or tragacanth, sodium starchglycolate, bentonite, a natural sponge, a surfactant, a resin such as acation-exchange resin, citrus pulp, sodium lauryl sulfate, sodium laurylsulfate in combination starch, and the like.

“Drug absorption” or “absorption” typically refers to the process ofmovement of drug from site of administration of a drug across a barrierinto a blood vessel or the site of action, e.g., a drug moving from thegastrointestinal tract into the portal vein or lymphatic system.

An “enteric coating” is a substance that remains substantially intact inthe stomach but dissolves and releases the drug in the small intestineor colon. Generally, the enteric coating comprises a polymeric materialthat prevents release in the low pH environment of the stomach but thationizes at a higher pH, typically a pH of 6 to 7, and thus dissolvessufficiently in the small intestine or colon to release the active agenttherein.

“Erosion facilitators” include materials that control the erosion of aparticular material in gastrointestinal fluid. Erosion facilitators aregenerally known to those of ordinary skill in the art. Exemplary erosionfacilitators include, e.g., hydrophilic polymers, electrolytes,proteins, peptides, and amino acids.

“Filling agents” include compounds such as lactose, calcium carbonate,calcium phosphate, dibasic calcium phosphate, calcium sulfate,microcrystalline cellulose, cellulose powder, dextrose, dextrates,dextran, starches, pregelatinized starch, sucrose, xylitol, lactitol,mannitol, sorbitol, sodium chloride, polyethylene glycol, and the like.

“Flavoring agents” and/or “sweeteners” useful in the formulationsdescribed herein, include, e.g., acacia syrup, acesulfame K, alitame,anise, apple, aspartame, banana, Bavarian cream, berry, black currant,butterscotch, calcium citrate, camphor, caramel, cherry, cherry cream,chocolate, cinnamon, bubble gum, citrus, citrus punch, citrus cream,cotton candy, cocoa, cola, cool cherry, cool citrus, cyclamate,cylamate, dextrose, eucalyptus, eugenol, fructose, fruit punch, ginger,glycyrrhetinate, glycyrrhiza (licorice) syrup, grape, grapefruit, honey,isomalt, lemon, lime, lemon cream, monoammonium glyrrhizinate(MagnaSweet®), maltol, mannitol, maple, marshmallow, menthol, mintcream, mixed berry, neohesperidine DC, neotame, orange, pear, peach,peppermint, peppermint cream, Prosweet® Powder, raspberry, root beer,rum, saccharin, safrole, sorbitol, spearmint, spearmint cream,strawberry, strawberry cream, stevia, sucralose, sucrose, sodiumsaccharin, saccharin, aspartame, acesulfame potassium, mannitol, talin,sylitol, sucralose, sorbitol, Swiss cream, tagatose, tangerine,thaumatin, tutti fruitti, vanilla, walnut, watermelon, wild cherry,wintergreen, xylitol, or any combination of these flavoring ingredients,e.g., anise-menthol, cherry-anise, cinnamon-orange, cherry-cinnamon,chocolate-mint, honey-lemon, lemon-lime, lemon-mint, menthol-eucalyptus,orange-cream, vanilla-mint, and mixtures thereof.

“Lubricants” and “glidants” are compounds that prevent, reduce orinhibit adhesion or friction of materials. Exemplary lubricants include,e.g., stearic acid, calcium hydroxide, talc, sodium stearyl fumerate, ahydrocarbon such as mineral oil, or hydrogenated vegetable oil such ashydrogenated soybean oil (Sterotex®), higher fatty acids and theiralkali-metal and alkaline earth metal salts, such as aluminum, calcium,magnesium, zinc, stearic acid, sodium stearates, glycerol, talc, waxes,Stearowet®, boric acid, sodium benzoate, sodium acetate, sodiumchloride, leucine, a polyethylene glycol (e.g., PEG-4000) or amethoxypolyethylene glycol such as Carbowax™, sodium oleate, sodiumbenzoate, glyceryl behenate, polyethylene glycol, magnesium or sodiumlauryl sulfate, colloidal silica such as Syloid™, Cab-O-Sil®, a starchsuch as corn starch, silicone oil, a surfactant, and the like.

A “measurable serum concentration” or “measurable plasma concentration”describes the blood serum or blood plasma concentration, typicallymeasured in mg, μg, or ng of therapeutic agent per ml, dl, or 1 of bloodserum, absorbed into the bloodstream after administration. As usedherein, measurable plasma concentrations are typically measured in ng/mlor μg/ml.

“Pharmacodynamics” refers to the factors which determine the biologicresponse observed relative to the concentration of drug at a site ofaction.

“Pharmacokinetics” refers to the factors which determine the attainmentand maintenance of the appropriate concentration of drug at a site ofaction.

“Plasticizers” are compounds used to soften the microencapsulationmaterial or film coatings to make them less brittle. Suitableplasticizers include, e.g., polyethylene glycols such as PEG 300, PEG400, PEG 600, PEG 1450, PEG 3350, and PEG 800, stearic acid, propyleneglycol, oleic acid, triethyl cellulose and triacetin. In someembodiments, plasticizers can also function as dispersing agents orwetting agents.

“Solubilizers” include compounds such as triacetin, triethylcitrate,ethyl oleate, ethyl caprylate, sodium lauryl sulfate, sodium doccusate,vitamin E TPGS, dimethylacetamide, N-methylpyrrolidone,N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropylmethylcellulose, hydroxypropyl cyclodextrins, ethanol, n-butanol, isopropylalcohol, cholesterol, bile salts, polyethylene glycol 200-600,glycofurol, transcutol, propylene glycol, and dimethyl isosorbide andthe like.

“Stabilizers” include compounds such as any antioxidation agents,buffers, acids, preservatives and the like.

“Steady state,” as used herein, is when the amount of drug administeredis equal to the amount of drug eliminated within one dosing intervalresulting in a plateau or constant plasma drug exposure.

“Suspending agents” include compounds such as polyvinylpyrrolidone,e.g., polyvinylpyrrolidone K12, polyvinylpyrrolidone K17,polyvinylpyrrolidone K25, or polyvinylpyrrolidone K30, vinylpyrrolidone/vinyl acetate copolymer (S630), polyethylene glycol, e.g.,the polyethylene glycol can have a molecular weight of about 300 toabout 6000, or about 3350 to about 4000, or about 7000 to about 5400,sodium carboxymethylcellulose, methylcellulose,hydroxypropylmethylcellulose, hydroxymethylcellulose acetate stearate,polysorbate-80, hydroxyethylcellulose, sodium alginate, gums, such as,e.g., gum tragacanth and gum acacia, guar gum, xanthans, includingxanthan gum, sugars, cellulosics, such as, e.g., sodiumcarboxymethylcellulose, methylcellulose, sodium carboxymethylcellulose,hydroxypropylmethylcellulose, hydroxyethylcellulose, polysorbate-80,sodium alginate, polyethoxylated sorbitan monolaurate, polyethoxylatedsorbitan monolaurate, povidone and the like.

“Surfactants” include compounds such as sodium lauryl sulfate, sodiumdocusate, Tween 60 or 80, triacetin, vitamin E TPGS, sorbitanmonooleate, polyoxyethylene sorbitan monooleate, polysorbates,polaxomers, bile salts, glyceryl monostearate, copolymers of ethyleneoxide and propylene oxide, e.g., Pluronic® (BASF), and the like. Someother surfactants include polyoxyethylene fatty acid glycerides andvegetable oils, e.g., polyoxyethylene (60) hydrogenated castor oil; andpolyoxyethylene alkylethers and alkylphenyl ethers, e.g., octoxynol 10,octoxynol 40. In some embodiments, surfactants may be included toenhance physical stability or for other purposes.

“Viscosity enhancing agents” include, e.g., methyl cellulose, xanthangum, carboxymethyl cellulose, hydroxypropyl cellulose,hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose acetatestearate, hydroxypropylmethyl cellulose phthalate, carbomer, polyvinylalcohol, alginates, acacia, chitosans and combinations thereof.

“Wetting agents” include compounds such as oleic acid, glycerylmonostearate, sorbitan monooleate, sorbitan monolaurate, triethanolamineoleate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitanmonolaurate, sodium docusate, sodium oleate, sodium lauryl sulfate,sodium doccusate, triacetin, Tween 80, vitamin E TPGS, ammonium saltsand the like.

Dosage Forms

The compositions described herein can be formulated for administrationto a subject via any conventional means including, but not limited to,oral, parenteral (e.g., intravenous, subcutaneous, or intramuscular),buccal, intranasal, rectal or transdermal administration routes. As usedherein, the term “subject” is used to mean an animal, preferably amammal, including a human or non-human. The terms patient and subjectmay be used interchangeably.

Moreover, the pharmaceutical compositions described herein, whichinclude a compound of any of Formula (I), (Ia)-(Ic), (IIa)-(IId),(IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId),(VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh)can be formulated into any suitable dosage form, including but notlimited to, aqueous oral dispersions, liquids, gels, syrups, elixirs,slurries, suspensions and the like, for oral ingestion by a patient tobe treated, solid oral dosage forms, aerosols, controlled releaseformulations, fast melt formulations, effervescent formulations,lyophilized formulations, tablets, powders, pills, dragees, capsules,delayed release formulations, extended release formulations, pulsatilerelease formulations, multiparticulate formulations, and mixed immediaterelease and controlled release formulations.

Pharmaceutical preparations for oral use can be obtained by mixing oneor more solid excipient with one or more of the compounds describedherein, optionally grinding the resulting mixture, and processing themixture of granules, after adding suitable auxiliaries, if desired, toobtain tablets or dragee cores. Suitable excipients include, forexample, fillers such as sugars, including lactose, sucrose, mannitol,or sorbitol; cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methylcellulose, microcrystalline cellulose,hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or otherssuch as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. Ifdesired, disintegrating agents may be added, such as the cross-linkedcroscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or asalt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers may be added. All formulations fororal administration should be in dosages suitable for suchadministration.

In some embodiments, the solid dosage forms disclosed herein may be inthe form of a tablet, (including a suspension tablet, a fast-melttablet, a bite-disintegration tablet, a rapid-disintegration tablet, aneffervescent tablet, or a caplet), a pill, a powder (including a sterilepackaged powder, a dispensable powder, or an effervescent powder) acapsule (including both soft or hard capsules, e.g., capsules made fromanimal-derived gelatin or plant-derived HPMC, or “sprinkle capsules”),solid dispersion, solid solution, bioerodible dosage form, controlledrelease formulations, pulsatile release dosage forms, multiparticulatedosage forms, pellets, granules, or an aerosol. In other embodiments,the pharmaceutical composition is in the form of a powder. In stillother embodiments, the pharmaceutical composition is in the form of atablet, including but not limited to, a fast-melt tablet. Additionally,pharmaceutical compositions described herein may be administered as asingle capsule or in multiple capsule dosage form. In some embodiments,the pharmaceutical composition is administered in two, or three, orfour, capsules or tablets.

In some embodiments, solid dosage forms, e.g., tablets, effervescenttablets, and capsules, are prepared by mixing particles of a compound ofany of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) with one or more pharmaceuticalexcipients to form a bulk blend composition. When referring to thesebulk blend compositions as homogeneous, it is meant that the particlesof the compound of any of Formula (I), (Ia)-(Ic), (IIa)-(IId),(IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId),(VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh)are dispersed evenly throughout the composition so that the compositionmay be readily subdivided into equally effective unit dosage forms, suchas tablets, pills, and capsules. The individual unit dosages may alsoinclude film coatings, which disintegrate upon oral ingestion or uponcontact with diluent. These formulations can be manufactured byconventional pharmacological techniques.

Conventional pharmacological techniques include, e.g., one or acombination of methods: (1) dry mixing, (2) direct compression, (3)milling, (4) dry or non-aqueous granulation, (5) wet granulation, or (6)fusion. See, e.g., Lachman et al., The Theory and Practice of IndustrialPharmacy (1986). Other methods include, e.g., spray drying, pan coating,melt granulation, granulation, fluidized bed spray drying or coating(e.g., wurster coating), tangential coating, top spraying, tableting,extruding and the like.

The pharmaceutical solid dosage forms described herein can include acompound described herein and one or more pharmaceutically acceptableadditives such as a compatible carrier, binder, filling agent,suspending agent, flavoring agent, sweetening agent, disintegratingagent, dispersing agent, surfactant, lubricant, colorant, diluent,solubilizer, moistening agent, plasticizer, stabilizer, penetrationenhancer, wetting agent, anti-foaming agent, antioxidant, preservative,or one or more combination thereof. In still other aspects, usingstandard coating procedures, such as those described in Remington'sPharmaceutical Sciences, 20th Edition (2000), a film coating is providedaround the formulation of the compound of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh). In one embodiment, some or all of the particles of thecompound of any of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) are coated. Inanother embodiment, some or all of the particles of the compound of anyof Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh), are microencapsulated. In stillanother embodiment, the particles of the compound of any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) are not microencapsulated and are uncoated.

Suitable carriers for use in the solid dosage forms described hereininclude, but are not limited to, acacia, gelatin, colloidal silicondioxide, calcium glycerophosphate, calcium lactate, maltodextrin,glycerine, magnesium silicate, sodium caseinate, soy lecithin, sodiumchloride, tricalcium phosphate, dipotassium phosphate, sodium stearoyllactylate, carrageenan, monoglyceride, diglyceride, pregelatinizedstarch, hydroxypropylmethylcellulose, hydroxypropylmethylcelluloseacetate stearate, sucrose, microcrystalline cellulose, lactose, mannitoland the like.

Suitable filling agents for use in the solid dosage forms describedherein include, but are not limited to, lactose, calcium carbonate,calcium phosphate, dibasic calcium phosphate, calcium sulfate,microcrystalline cellulose, cellulose powder, dextrose, dextrates,dextran, starches, pregelatinized starch, hydroxypropylmethycellulose(HPMC), hydroxypropylmethycellulose phthalate,hydroxypropylmethylcellulose acetate stearate (HPMCAS), sucrose,xylitol, lactitol, mannitol, sorbitol, sodium chloride, polyethyleneglycol, and the like.

In order to release the compound of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) from a solid dosage form matrix as efficiently aspossible, disintegrants are often used in the formulation, especiallywhen the dosage forms are compressed with binder. Disintegrants helprupturing the dosage form matrix by swelling or capillary action whenmoisture is absorbed into the dosage form. Suitable disintegrants foruse in the solid dosage forms described herein include, but are notlimited to, natural starch such as corn starch or potato starch, apregelatinized starch such as National 1551 or Amijel®, or sodium starchglycolate such as Promogel® or Explotab®, a cellulose such as a woodproduct, methylcrystalline cellulose, e.g., Avicel®, Avicel® PH101,Avicel® PH102, Avicel® PH105, Elcema® P100, Emcocel®, Vivacel®, MingTia®, and Solka-Floc®, methylcellulose, croscarmellose, or across-linked cellulose, such as cross-linked sodiumcarboxymethylcellulose (Ac-Di-Sol®), cross-linkedcarboxymethylcellulose, or cross-linked croscarmellose, a cross-linkedstarch such as sodium starch glycolate, a cross-linked polymer such ascrospovidone, a cross-linked polyvinylpyrrolidone, alginate such asalginic acid or a salt of alginic acid such as sodium alginate, a claysuch as Veegum® HV (magnesium aluminum silicate), a gum such as agar,guar, locust bean, Karaya, pectin, or tragacanth, sodium starchglycolate, bentonite, a natural sponge, a surfactant, a resin such as acation-exchange resin, citrus pulp, sodium lauryl sulfate, sodium laurylsulfate in combination starch, and the like.

Binders impart cohesiveness to solid oral dosage form formulations: forpowder filled capsule formulation, they aid in plug formation that canbe filled into soft or hard shell capsules and for tablet formulation,they ensure the tablet remaining intact after compression and helpassure blend uniformity prior to a compression or fill step. Materialssuitable for use as binders in the solid dosage forms described hereininclude, but are not limited to, carboxymethylcellulose, methylcellulose(e.g., Methocel®), hydroxypropylmethylcellulose (e.g. Hypromellose USPPharmacoat-603, hydroxypropylmethylcellulose acetate stearate (AqoateHS-LF and HS), hydroxyethylcellulose, hydroxypropylcellulose (e.g.,Klucel®), ethylcellulose (e.g., Ethocel®), and microcrystallinecellulose (e.g., Avicel®), microcrystalline dextrose, amylose, magnesiumaluminum silicate, polysaccharide acids, bentonites, gelatin,polyvinylpyrrolidone/vinyl acetate copolymer, crospovidone, povidone,starch, pregelatinized starch, tragacanth, dextrin, a sugar, such assucrose (e.g., Dipac®), glucose, dextrose, molasses, mannitol, sorbitol,xylitol (e.g., Xylitab®), lactose, a natural or synthetic gum such asacacia, tragacanth, ghatti gum, mucilage of isapol husks, starch,polyvinylpyrrolidone (e.g., Povidone® CL, Kollidon® CL, Polyplasdone®XL-10, and Povidone® K-12), larch arabogalactan, Veegum®, polyethyleneglycol, waxes, sodium alginate, and the like.

In general, binder levels of 20-70% are used in powder-filled gelatincapsule formulations. Binder usage level in tablet formulations varieswhether direct compression, wet granulation, roller compaction, or usageof other excipients such as fillers which itself can act as moderatebinder. Formulators skilled in art can determine the binder level forthe formulations, but binder usage level of up to 70% in tabletformulations is common.

Suitable lubricants or glidants for use in the solid dosage formsdescribed herein include, but are not limited to, stearic acid, calciumhydroxide, talc, corn starch, sodium stearyl fumerate, alkali-metal andalkaline earth metal salts, such as aluminum, calcium, magnesium, zinc,stearic acid, sodium stearates, magnesium stearate, zinc stearate,waxes, Stearowet®, boric acid, sodium benzoate, sodium acetate, sodiumchloride, leucine, a polyethylene glycol or a methoxypolyethylene glycolsuch as Carbowax™, PEG 4000, PEG 5000, PEG 6000, propylene glycol,sodium oleate, glyceryl behenate, glyceryl palmitostearate, glycerylbenzoate, magnesium or sodium lauryl sulfate, and the like.

Suitable diluents for use in the solid dosage forms described hereininclude, but are not limited to, sugars (including lactose, sucrose, anddextrose), polysaccharides (including dextrates and maltodextrin),polyols (including mannitol, xylitol, and sorbitol), cyclodextrins andthe like.

The term “non water-soluble diluent” represents compounds typically usedin the formulation of pharmaceuticals, such as calcium phosphate,calcium sulfate, starches, modified starches and microcrystallinecellulose, and microcellulose (e.g., having a density of about 0.45g/cm³, e.g. Avicel, powdered cellulose), and talc.

Suitable wetting agents for use in the solid dosage forms describedherein include, for example, oleic acid, glyceryl monostearate, sorbitanmonooleate, sorbitan monolaurate, triethanolamine oleate,polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitanmonolaurate, quaternary ammonium compounds (e.g., Polyquat 10®), sodiumoleate, sodium lauryl sulfate, magnesium stearate, sodium docusate,triacetin, vitamin E TPGS and the like.

Suitable surfactants for use in the solid dosage forms described hereininclude, for example, sodium lauryl sulfate, sorbitan monooleate,polyoxyethylene sorbitan monooleate, polysorbates, polaxomers, bilesalts, glyceryl monostearate, copolymers of ethylene oxide and propyleneoxide, e.g., Pluronic® (BASF), and the like.

Suitable suspending agents for use in the solid dosage forms describedhere include, but are not limited to, polyvinylpyrrolidone, e.g.,polyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidoneK25, or polyvinylpyrrolidone K30, polyethylene glycol, e.g., thepolyethylene glycol can have a molecular weight of about 300 to about6000, or about 3350 to about 4000, or about 7000 to about 5400, vinylpyrrolidone/vinyl acetate copolymer (S630), sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,polysorbate-80, hydroxyethylcellulose, sodium alginate, gums, such as,e.g., gum tragacanth and gum acacia, guar gum, xanthans, includingxanthan gum, sugars, cellulosics, such as, e.g., sodiumcarboxymethylcellulose, methylcellulose, sodium carboxymethylcellulose,hydroxypropylmethylcellulose, hydroxyethylcellulose, polysorbate-80,sodium alginate, polyethoxylated sorbitan monolaurate, polyethoxylatedsorbitan monolaurate, povidone and the like.

Suitable antioxidants for use in the solid dosage forms described hereininclude, for example, e.g., butylated hydroxytoluene (BHT), sodiumascorbate, and tocopherol.

It should be appreciated that there is considerable overlap betweenadditives used in the solid dosage forms described herein. Thus, theabove-listed additives should be taken as merely exemplary, and notlimiting, of the types of additives that can be included in solid dosageforms described herein. The amounts of such additives can be readilydetermined by one skilled in the art, according to the particularproperties desired.

In other embodiments, one or more layers of the pharmaceuticalcomposition are plasticized. Illustratively, a plasticizer is generallya high boiling point solid or liquid. Suitable plasticizers can be addedfrom about 0.01% to about 50% by weight (w/w) of the coatingcomposition. Plasticizers include, but are not limited to, diethylphthalate, citrate esters, polyethylene glycol, glycerol, acetylatedglycerides, triacetin, polypropylene glycol, polyethylene glycol,triethyl citrate, dibutyl sebacate, stearic acid, stearol, stearate, andcastor oil.

Compressed tablets are solid dosage forms prepared by compacting thebulk blend of the formulations described above. In various embodiments,compressed tablets which are designed to dissolve in the mouth willinclude one or more flavoring agents. In other embodiments, thecompressed tablets will include a film surrounding the final compressedtablet. In some embodiments, the film coating can provide a delayedrelease of the compound of any of Formula (I), (Ia)-(Ic), (IIa)-(IId),(IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId),(VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh)from the formulation. In other embodiments, the film coating aids inpatient compliance (e.g., Opadry® coatings or sugar coating). Filmcoatings including Opadry® typically range from about 1% to about 3% ofthe tablet weight. In other embodiments, the compressed tablets includeone or more excipients.

A capsule may be prepared, for example, by placing the bulk blend of theformulation of the compound of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh), described above, inside of a capsule. In someembodiments, the formulations (non-aqueous suspensions and solutions)are placed in a soft gelatin capsule. In other embodiments, theformulations are placed in standard gelatin capsules or non-gelatincapsules such as capsules comprising HPMC. In other embodiments, theformulation is placed in a sprinkle capsule, wherein the capsule may beswallowed whole or the capsule may be opened and the contents sprinkledon food prior to eating. In some embodiments, the therapeutic dose issplit into multiple (e.g., two, three, or four) capsules. In someembodiments, the entire dose of the formulation is delivered in acapsule form.

In various embodiments, the particles of the compound of any of Formula(I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) and one or more excipients are dry blendedand compressed into a mass, such as a tablet, having a hardnesssufficient to provide a pharmaceutical composition that substantiallydisintegrates within less than about 30 minutes, less than about 35minutes, less than about 40 minutes, less than about 45 minutes, lessthan about 50 minutes, less than about 55 minutes, or less than about 60minutes, after oral administration, thereby releasing the formulationinto the gastrointestinal fluid.

In another aspect, dosage forms may include microencapsulatedformulations. In some embodiments, one or more other compatiblematerials are present in the microencapsulation material. Exemplarymaterials include, but are not limited to, pH modifiers, erosionfacilitators, anti-foaming agents, antioxidants, flavoring agents, andcarrier materials such as binders, suspending agents, disintegrationagents, filling agents, surfactants, solubilizers, stabilizers,lubricants, wetting agents, and diluents.

Materials useful for the microencapsulation described herein includematerials compatible with compounds of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) which sufficiently isolate the compound of any of Formula(I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) from other non-compatible excipients.Materials compatible with compounds of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) are those that delay the release of the compounds of anyof Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh), in vivo.

Exemplary microencapsulation materials useful for delaying the releaseof the formulations including compounds described herein, include, butare not limited to, hydroxypropyl cellulose ethers (HPC) such as Klucel®or Nisso HPC, low-substituted hydroxypropyl cellulose ethers (L-HPC),hydroxypropyl methyl cellulose ethers (HPMC) such as Seppifilm-LC,Pharmacoat®, Metolose SR, Methocel®-E, Opadry YS, PrimaFlo, BenecelMP824, and Benecel MP843, methylcellulose polymers such as Methocel®-A,hydroxypropylmethylcellulose acetate stearate Aqoat (HF-LS, HF-LG,HF-MS) and Metolose®, Ethylcelluloses (EC) and mixtures thereof such asE461, Ethocel®, Aqualon®-EC, Surelease®, Polyvinyl alcohol (PVA) such asOpadry AMB, hydroxyethylcelluloses such as Natrosol®,carboxymethylcelluloses and salts of carboxymethylcelluloses (CMC) suchas Aqualon®-CMC, polyvinyl alcohol and polyethylene glycol co-polymerssuch as Kollicoat IR®, monoglycerides (Myverol), triglycerides (KLX),polyethylene glycols, modified food starch, acrylic polymers andmixtures of acrylic polymers with cellulose ethers such as Eudragit®EPO, Eudragit® L30D-55, Eudragit® FS 30D Eudragit® L100-55, Eudragit®L100, Eudragit® S100, Eudragit® RD100, Eudragit® E100, Eudragit® L12.5,Eudragit® S12.5, Eudragit® NE30D, and Eudragit® NE 40D, celluloseacetate phthalate, sepifilms such as mixtures of HPMC and stearic acid,cyclodextrins, and mixtures of these materials.

In still other embodiments, plasticizers such as polyethylene glycols,e.g., PEG 300, PEG 400, PEG 600, PEG 1450, PEG 3350, and PEG 800,stearic acid, propylene glycol, oleic acid, and triacetin areincorporated into the microencapsulation material. In other embodiments,the microencapsulating material useful for delaying the release of thepharmaceutical compositions is from the USP or the National Formulary(NF). In yet other embodiments, the microencapsulation material isKlucel. In still other embodiments, the microencapsulation material ismethocel.

Microencapsulated compounds of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) may be formulated by methods known by one of ordinaryskill in the art. Such known methods include, e.g., spray dryingprocesses, spinning disk-solvent processes, hot melt processes, spraychilling methods, fluidized bed, electrostatic deposition, centrifugalextrusion, rotational suspension separation, polymerization atliquid-gas or solid-gas interface, pressure extrusion, or sprayingsolvent extraction bath. In addition to these, several chemicaltechniques, e.g., complex coacervation, solvent evaporation,polymer-polymer incompatibility, interfacial polymerization in liquidmedia, in situ polymerization, in-liquid drying, and desolvation inliquid media could also be used. Furthermore, other methods such asroller compaction, extrusion/spheronization, coacervation, ornanoparticle coating may also be used.

In one embodiment, the particles of compounds of any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) are microencapsulated prior to beingformulated into one of the above forms. In still another embodiment,some or most of the particles are coated prior to being furtherformulated by using standard coating procedures, such as those describedin Remington's Pharmaceutical Sciences, 20th Edition (2000).

In other embodiments, the solid dosage formulations of the compounds ofany of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) are plasticized (coated) withone or more layers. Illustratively, a plasticizer is generally a highboiling point solid or liquid. Suitable plasticizers can be added fromabout 0.01% to about 50% by weight (w/w) of the coating composition.Plasticizers include, but are not limited to, diethyl phthalate, citrateesters, polyethylene glycol, glycerol, acetylated glycerides, triacetin,polypropylene glycol, polyethylene glycol, triethyl citrate, dibutylsebacate, stearic acid, stearol, stearate, and castor oil.

In other embodiments, a powder including the formulations with acompound of any of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh), described herein,may be formulated to include one or more pharmaceutical excipients andflavors. Such a powder may be prepared, for example, by mixing theformulation and optional pharmaceutical excipients to form a bulk blendcomposition. Additional embodiments also include a suspending agentand/or a wetting agent. This bulk blend is uniformly subdivided intounit dosage packaging or multi-dosage packaging units.

In still other embodiments, effervescent powders are also prepared inaccordance with the present disclosure. Effervescent salts have beenused to disperse medicines in water for oral administration.Effervescent salts are granules or coarse powders containing a medicinalagent in a dry mixture, usually composed of sodium bicarbonate, citricacid and/or tartaric acid. When salts of the compositions describedherein are added to water, the acids and the base react to liberatecarbon dioxide gas, thereby causing “effervescence.” Examples ofeffervescent salts include, e.g., the following ingredients: sodiumbicarbonate or a mixture of sodium bicarbonate and sodium carbonate,citric acid and/or tartaric acid. Any acid-base combination that resultsin the liberation of carbon dioxide can be used in place of thecombination of sodium bicarbonate and citric and tartaric acids, as longas the ingredients were suitable for pharmaceutical use and result in apH of about 6.0 or higher.

In other embodiments, the formulations described herein, which include acompound of Formula (A), are solid dispersions. Methods of producingsuch solid dispersions are known in the art and include, but are notlimited to, for example, U.S. Pat. Nos. 4,343,789, 5,340,591, 5,456,923,5,700,485, 5,723,269, and U.S. Pub. Appl 2004/0013734, each of which isspecifically incorporated by reference. In still other embodiments, theformulations described herein are solid solutions. Solid solutionsincorporate a substance together with the active agent and otherexcipients such that heating the mixture results in dissolution of thedrug and the resulting composition is then cooled to provide a solidblend which can be further formulated or directly added to a capsule orcompressed into a tablet. Methods of producing such solid solutions areknown in the art and include, but are not limited to, for example, U.S.Pat. Nos. 4,151,273, 5,281,420, and 6,083,518, each of which isspecifically incorporated by reference.

The pharmaceutical solid oral dosage forms including formulationsdescribed herein, which include a compound of any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) can be further formulated to provide acontrolled release of the compound of Formula (A). Controlled releaserefers to the release of the compound of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) from a dosage form in which it is incorporated accordingto a desired profile over an extended period of time. Controlled releaseprofiles include, for example, sustained release, prolonged release,pulsatile release, and delayed release profiles. In contrast toimmediate release compositions, controlled release compositions allowdelivery of an agent to a subject over an extended period of timeaccording to a predetermined profile. Such release rates can providetherapeutically effective levels of agent for an extended period of timeand thereby provide a longer period of pharmacologic response whileminimizing side effects as compared to conventional rapid release dosageforms. Such longer periods of response provide for many inherentbenefits that are not achieved with the corresponding short acting,immediate release preparations.

In some embodiments, the solid dosage forms described herein can beformulated as enteric coated delayed release oral dosage forms, i.e., asan oral dosage form of a pharmaceutical composition as described hereinwhich utilizes an enteric coating to affect release in the smallintestine of the gastrointestinal tract. The enteric coated dosage formmay be a compressed or molded or extruded tablet/mold (coated oruncoated) containing granules, powder, pellets, beads or particles ofthe active ingredient and/or other composition components, which arethemselves coated or uncoated. The enteric coated oral dosage form mayalso be a capsule (coated or uncoated) containing pellets, beads orgranules of the solid carrier or the composition, which are themselvescoated or uncoated.

The term “delayed release” as used herein refers to the delivery so thatthe release can be accomplished at some generally predictable locationin the intestinal tract more distal to that which would have beenaccomplished if there had been no delayed release alterations. In someembodiments the method for delay of release is coating. Any coatingsshould be applied to a sufficient thickness such that the entire coatingdoes not dissolve in the gastrointestinal fluids at pH below about 5,but does dissolve at pH about 5 and above. It is expected that anyanionic polymer exhibiting a pH-dependent solubility profile can be usedas an enteric coating in the methods and compositions described hereinto achieve delivery to the lower gastrointestinal tract. In someembodiments the polymers described herein are anionic carboxylicpolymers. In other embodiments, the polymers and compatible mixturesthereof, and some of their properties, include, but are not limited to:

Shellac, also called purified lac, a refined product obtained from theresinous secretion of an insect. This coating dissolves in media ofpH>7;

Acrylic polymers. The performance of acrylic polymers (primarily theirsolubility in biological fluids) can vary based on the degree and typeof substitution. Examples of suitable acrylic polymers includemethacrylic acid copolymers and ammonium methacrylate copolymers. TheEudragit series E, L, S, RL, RS and NE (Rohm Pharma) are available assolubilized in organic solvent, aqueous dispersion, or dry powders. TheEudragit series RL, NE, and RS are insoluble in the gastrointestinaltract but are permeable and are used primarily for colonic targeting.The Eudragit series E dissolve in the stomach. The Eudragit series L,L-30D and S are insoluble in stomach and dissolve in the intestine;

Cellulose Derivatives. Examples of suitable cellulose derivatives are:ethyl cellulose; reaction mixtures of partial acetate esters ofcellulose with phthalic anhydride. The performance can vary based on thedegree and type of substitution. Cellulose acetate phthalate (CAP)dissolves in pH>6. Aquateric (FMC) is an aqueous based system and is aspray dried CAP psuedolatex with particles <1 μm. Other components inAquateric can include pluronics, Tweens, and acetylated monoglycerides.Other suitable cellulose derivatives include: cellulose acetatetrimellitate (Eastman); methylcellulose (Pharmacoat, Methocel);hydroxypropylmethyl cellulose phthalate (HPMCP); hydroxypropylmethylcellulose succinate (HPMCS); and hydroxypropylmethylcellulose acetatesuccinate (e.g., AQOAT (Shin Etsu)). The performance can vary based onthe degree and type of substitution. For example, HPMCP such as, HP-50,HP-55, HP-555, HP-55F grades are suitable. The performance can varybased on the degree and type of substitution. For example, suitablegrades of hydroxypropylmethylcellulose acetate succinate include, butare not limited to, AS-LG (LF), which dissolves at pH 5, AS-MG (MF),which dissolves at pH 5.5, and AS-HG (HF), which dissolves at higher pH.These polymers are offered as granules, or as fine powders for aqueousdispersions;

Poly Vinyl Acetate Phthalate (PVAP). PVAP dissolves in pH>5, and it ismuch less permeable to water vapor and gastric fluids.

In some embodiments, the coating can, and usually does, contain aplasticizer and possibly other coating excipients such as colorants,talc, and/or magnesium stearate, which are well known in the art.Suitable plasticizers include triethyl citrate (Citroflex 2), triacetin(glyceryl triacetate), acetyl triethyl citrate (Citroflec A2), Carbowax400 (polyethylene glycol 400), diethyl phthalate, tributyl citrate,acetylated monoglycerides, glycerol, fatty acid esters, propyleneglycol, and dibutyl phthalate. In particular, anionic carboxylic acrylicpolymers usually will contain 10-25% by weight of a plasticizer,especially dibutyl phthalate, polyethylene glycol, triethyl citrate andtriacetin. Conventional coating techniques such as spray or pan coatingare employed to apply coatings. The coating thickness must be sufficientto ensure that the oral dosage form remains intact until the desiredsite of topical delivery in the intestinal tract is reached.

Colorants, detackifiers, surfactants, antifoaming agents, lubricants(e.g., carnuba wax or PEG) may be added to the coatings besidesplasticizers to solubilize or disperse the coating material, and toimprove coating performance and the coated product.

In other embodiments, the formulations described herein, which include acompound of Formula (A), are delivered using a pulsatile dosage form. Apulsatile dosage form is capable of providing one or more immediaterelease pulses at predetermined time points after a controlled lag timeor at specific sites. Pulsatile dosage forms including the formulationsdescribed herein, which include a compound of any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) may be administered using a variety ofpulsatile formulations known in the art. For example, such formulationsinclude, but are not limited to, those described in U.S. Pat. Nos.5,011,692, 5,017,381, 5,229,135, and 5,840,329, each of which isspecifically incorporated by reference. Other pulsatile release dosageforms suitable for use with the present formulations include, but arenot limited to, for example, U.S. Pat. Nos. 4,871,549, 5,260,068,5,260,069, 5,508,040, 5,567,441 and 5,837,284, all of which arespecifically incorporated by reference. In one embodiment, thecontrolled release dosage form is pulsatile release solid oral dosageform including at least two groups of particles, (i.e. multiparticulate)each containing the formulation described herein. The first group ofparticles provides a substantially immediate dose of the compound of anyof Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) upon ingestion by a mammal. Thefirst group of particles can be either uncoated or include a coatingand/or sealant. The second group of particles includes coated particles,which includes from about 2% to about 75%, from about 2.5% to about 70%,or from about 40% to about 70%, by weight of the total dose of thecompound of any of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) in saidformulation, in admixture with one or more binders. The coating includesa pharmaceutically acceptable ingredient in an amount sufficient toprovide a delay of from about 2 hours to about 7 hours followingingestion before release of the second dose. Suitable coatings includeone or more differentially degradable coatings such as, by way ofexample only, pH sensitive coatings (enteric coatings) such as acrylicresins (e.g., Eudragit® EPO, Eudragit® L30D-55, Eudragit® FS 30DEudragit® L100-55, Eudragit® L100, Eudragit® S100, Eudragit® RD100,Eudragit® E100, Eudragit® L12.5, Eudragit® S12.5, and Eudragit® NE30D,Eudragit® NE 40D®) either alone or blended with cellulose derivatives,e.g., ethylcellulose, or non-enteric coatings having variable thicknessto provide differential release of the formulation that includes acompound of any of Formula (I).

Many other types of controlled release systems known to those ofordinary skill in the art and are suitable for use with the formulationsdescribed herein. Examples of such delivery systems include, e.g.,polymer-based systems, such as polylactic and polyglycolic acid,plyanhydrides and polycaprolactone; porous matrices, nonpolymer-basedsystems that are lipids, including sterols, such as cholesterol,cholesterol esters and fatty acids, or neutral fats, such as mono-, di-and triglycerides; hydrogel release systems; silastic systems;peptide-based systems; wax coatings, bioerodible dosage forms,compressed tablets using conventional binders and the like. See, e.g.,Liberman et al., Pharmaceutical Dosage Forms, 2 Ed., Vol. 1, pp. 209-214(1990); Singh et al., Encyclopedia of Pharmaceutical Technology, 2^(nd)Ed., pp. 751-753 (2002); U.S. Pat. Nos. 4,327,725, 4,624,848, 4,968,509,5,461,140, 5,456,923, 5,516,527, 5,622,721, 5,686,105, 5,700,410,5,977,175, 6,465,014 and 6,932,983, each of which is specificallyincorporated by reference.

In some embodiments, pharmaceutical compositions are provided thatinclude particles of the compounds of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh), described herein and at least one dispersing agent orsuspending agent for oral administration to a subject. The formulationsmay be a powder and/or granules for suspension, and upon admixture withwater, a substantially uniform suspension is obtained.

Liquid formulation dosage forms for oral administration can be aqueoussuspensions selected from the group including, but not limited to,pharmaceutically acceptable aqueous oral dispersions, emulsions,solutions, elixirs, gels, and syrups. See, e.g., Singh et al.,Encyclopedia of Pharmaceutical Technology, 2^(nd) Ed., pp. 754-757(2002). In addition to the particles of compound of Formula (A), theliquid dosage forms may include additives, such as: (a) disintegratingagents; (b) dispersing agents; (c) wetting agents; (d) at least onepreservative, (e) viscosity enhancing agents, (f) at least onesweetening agent, and (g) at least one flavoring agent. In someembodiments, the aqueous dispersions can further include a crystallineinhibitor.

The aqueous suspensions and dispersions described herein can remain in ahomogenous state, as defined in The USP Pharmacists' Pharmacopeia (2005edition, chapter 905), for at least 4 hours. The homogeneity should bedetermined by a sampling method consistent with regard to determininghomogeneity of the entire composition. In one embodiment, an aqueoussuspension can be re-suspended into a homogenous suspension by physicalagitation lasting less than 1 minute. In another embodiment, an aqueoussuspension can be re-suspended into a homogenous suspension by physicalagitation lasting less than 45 seconds. In yet another embodiment, anaqueous suspension can be re-suspended into a homogenous suspension byphysical agitation lasting less than 30 seconds. In still anotherembodiment, no agitation is necessary to maintain a homogeneous aqueousdispersion.

Examples of disintegrating agents for use in the aqueous suspensions anddispersions include, but are not limited to, a starch, e.g., a naturalstarch such as corn starch or potato starch, a pregelatinized starchsuch as National 1551 or Amijel®, or sodium starch glycolate such asPromogel® or Explotab®; a cellulose such as a wood product,methylcrystalline cellulose, e.g., Avicel®, Avicel® PH101, Avicel®PH102, Avicel® PH105, Elcema® P100, Emcocel®, Vivacel®, Ming Tia®, andSolka-Floc®, methylcellulose, croscarmellose, or a cross-linkedcellulose, such as cross-linked sodium carboxymethylcellulose(Ac-Di-Sol®), cross-linked carboxymethylcellulose, or cross-linkedcroscarmellose; a cross-linked starch such as sodium starch glycolate; across-linked polymer such as crospovidone; a cross-linkedpolyvinylpyrrolidone; alginate such as alginic acid or a salt of alginicacid such as sodium alginate; a clay such as Veegum® HV (magnesiumaluminum silicate); a gum such as agar, guar, locust bean, Karaya,pectin, or tragacanth; sodium starch glycolate; bentonite; a naturalsponge; a surfactant; a resin such as a cation-exchange resin; citruspulp; sodium lauryl sulfate; sodium lauryl sulfate in combinationstarch; and the like.

In some embodiments, the dispersing agents suitable for the aqueoussuspensions and dispersions described herein are known in the art andinclude, for example, hydrophilic polymers, electrolytes, Tween® 60 or80, PEG, polyvinylpyrrolidone (PVP; commercially known as Plasdone®),and the carbohydrate-based dispersing agents such as, for example,hydroxypropylcellulose and hydroxypropyl cellulose ethers (e.g., HPC,HPC-SL, and HPC-L), hydroxypropyl methylcellulose and hydroxypropylmethylcellulose ethers (e.g. HPMC K100, HPMC K4M, HPMC K15M, and HPMCK100M), carboxymethylcellulose sodium, methylcellulose,hydroxyethylcellulose, hydroxypropylmethyl-cellulose phthalate,hydroxypropylmethyl-cellulose acetate stearate, noncrystallinecellulose, magnesium aluminum silicate, triethanolamine, polyvinylalcohol (PVA), polyvinylpyrrolidone/vinyl acetate copolymer (Plasdone®,e.g., S-630), 4-(1,1,3,3-tetramethylbutyl)-phenol polymer with ethyleneoxide and formaldehyde (also known as tyloxapol), poloxamers (e.g.,Pluronics F68®, F88®, and F108®, which are block copolymers of ethyleneoxide and propylene oxide); and poloxamines (e.g., Tetronic 908®, alsoknown as Poloxamine 908®, which is a tetrafunctional block copolymerderived from sequential addition of propylene oxide and ethylene oxideto ethylenediamine (BASF Corporation, Parsippany, N.J.)). In otherembodiments, the dispersing agent is selected from a group notcomprising one of the following agents: hydrophilic polymers;electrolytes; Tween® 60 or 80; PEG; polyvinylpyrrolidone (PVP);hydroxypropylcellulose and hydroxypropyl cellulose ethers (e.g., HPC,HPC-SL, and HPC-L); hydroxypropyl methylcellulose and hydroxypropylmethylcellulose ethers (e.g. HPMC K100, HPMC K4M, HPMC K15M, HPMC K100M,and Pharmacoat® USP 2910 (Shin-Etsu)); carboxymethylcellulose sodium;methylcellulose; hydroxyethylcellulose; hydroxypropylmethyl-cellulosephthalate; hydroxypropylmethyl-cellulose acetate stearate;non-crystalline cellulose; magnesium aluminum silicate; triethanolamine;polyvinyl alcohol (PVA); 4-(1,1,3,3-tetramethylbutyl)-phenol polymerwith ethylene oxide and formaldehyde; poloxamers (e.g., Pluronics F68®,F88®, and F108®, which are block copolymers of ethylene oxide andpropylene oxide); or poloxamines (e.g., Tetronic 908®, also known asPoloxamine 908®).

Wetting agents suitable for the aqueous suspensions and dispersionsdescribed herein are known in the art and include, but are not limitedto, cetyl alcohol, glycerol monostearate, polyoxyethylene sorbitan fattyacid esters (e.g., the commercially available Tweens® such as e.g.,Tween 20® and Tween 80® (ICI Specialty Chemicals)), and polyethyleneglycols (e.g., Carbowaxs 3350® and 1450®, and Carbopol 934® (UnionCarbide)), oleic acid, glyceryl monostearate, sorbitan monooleate,sorbitan monolaurate, triethanolamine oleate, polyoxyethylene sorbitanmonooleate, polyoxyethylene sorbitan monolaurate, sodium oleate, sodiumlauryl sulfate, sodium docusate, triacetin, vitamin E TPGS, sodiumtaurocholate, simethicone, phosphotidylcholine and the like

Suitable preservatives for the aqueous suspensions or dispersionsdescribed herein include, for example, potassium sorbate, parabens(e.g., methylparaben and propylparaben), benzoic acid and its salts,other esters of parahydroxybenzoic acid such as butylparaben, alcoholssuch as ethyl alcohol or benzyl alcohol, phenolic compounds such asphenol, or quaternary compounds such as benzalkonium chloride.Preservatives, as used herein, are incorporated into the dosage form ata concentration sufficient to inhibit microbial growth.

Suitable viscosity enhancing agents for the aqueous suspensions ordispersions described herein include, but are not limited to, methylcellulose, xanthan gum, carboxymethyl cellulose, hydroxypropylcellulose, hydroxypropylmethyl cellulose, Plasdon® S-630, carbomer,polyvinyl alcohol, alginates, acacia, chitosans and combinationsthereof. The concentration of the viscosity enhancing agent will dependupon the agent selected and the viscosity desired.

Examples of sweetening agents suitable for the aqueous suspensions ordispersions described herein include, for example, acacia syrup,acesulfame K, alitame, anise, apple, aspartame, banana, Bavarian cream,berry, black currant, butterscotch, calcium citrate, camphor, caramel,cherry, cherry cream, chocolate, cinnamon, bubble gum, citrus, citruspunch, citrus cream, cotton candy, cocoa, cola, cool cherry, coolcitrus, cyclamate, cylamate, dextrose, eucalyptus, eugenol, fructose,fruit punch, ginger, glycyrrhetinate, glycyrrhiza (licorice) syrup,grape, grapefruit, honey, isomalt, lemon, lime, lemon cream,monoammonium glyrrhizinate (MagnaSweet®), maltol, mannitol, maple,marshmallow, menthol, mint cream, mixed berry, neohesperidine DC,neotame, orange, pear, peach, peppermint, peppermint cream, Prosweet®Powder, raspberry, root beer, rum, saccharin, safrole, sorbitol,spearmint, spearmint cream, strawberry, strawberry cream, stevia,sucralose, sucrose, sodium saccharin, saccharin, aspartame, acesulfamepotassium, mannitol, talin, sucralose, sorbitol, swiss cream, tagatose,tangerine, thaumatin, tutti fruitti, vanilla, walnut, watermelon, wildcherry, wintergreen, xylitol, or any combination of these flavoringingredients, e.g., anise-menthol, cherry-anise, cinnamon-orange,cherry-cinnamon, chocolate-mint, honey-lemon, lemon-lime, lemon-mint,menthol-eucalyptus, orange-cream, vanilla-mint, and mixtures thereof. Inone embodiment, the aqueous liquid dispersion can comprise a sweeteningagent or flavoring agent in a concentration ranging from about 0.001% toabout 1.0% the volume of the aqueous dispersion. In another embodiment,the aqueous liquid dispersion can comprise a sweetening agent orflavoring agent in a concentration ranging from about 0.005% to about0.5% the volume of the aqueous dispersion. In yet another embodiment,the aqueous liquid dispersion can comprise a sweetening agent orflavoring agent in a concentration ranging from about 0.01% to about1.0% the volume of the aqueous dispersion.

In addition to the additives listed above, the liquid formulations canalso include inert diluents commonly used in the art, such as water orother solvents, solubilizing agents, and emulsifiers. Exemplaryemulsifiers are ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propyleneglycol,1,3-butyleneglycol, dimethylformamide, sodium lauryl sulfate, sodiumdoccusate, cholesterol, cholesterol esters, taurocholic acid,phosphotidylcholine, oils, such as cottonseed oil, groundnut oil, corngerm oil, olive oil, castor oil, and sesame oil, glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols, fatty acid esters ofsorbitan, or mixtures of these substances, and the like.

In some embodiments, the pharmaceutical compositions described hereincan be self-emulsifying drug delivery systems (SEDDS). Emulsions aredispersions of one immiscible phase in another, usually in the form ofdroplets. Generally, emulsions are created by vigorous mechanicaldispersion. SEDDS, as opposed to emulsions or microemulsions,spontaneously form emulsions when added to an excess of water withoutany external mechanical dispersion or agitation. An advantage of SEDDSis that only gentle mixing is required to distribute the dropletsthroughout the solution. Additionally, water or the aqueous phase can beadded just prior to administration, which ensures stability of anunstable or hydrophobic active ingredient. Thus, the SEDDS provides aneffective delivery system for oral and parenteral delivery ofhydrophobic active ingredients. SEDDS may provide improvements in thebioavailability of hydrophobic active ingredients. Methods of producingself-emulsifying dosage forms are known in the art and include, but arenot limited to, for example, U.S. Pat. Nos. 5,858,401, 6,667,048, and6,960,563, each of which is specifically incorporated by reference.

It is to be appreciated that there is overlap between the above-listedadditives used in the aqueous dispersions or suspensions describedherein, since a given additive is often classified differently bydifferent practitioners in the field, or is commonly used for any ofseveral different functions. Thus, the above-listed additives should betaken as merely exemplary, and not limiting, of the types of additivesthat can be included in formulations described herein. The amounts ofsuch additives can be readily determined by one skilled in the art,according to the particular properties desired.

Intranasal Formulations

Intranasal formulations are known in the art and are described in, forexample, U.S. Pat. Nos. 4,476,116, 5,116,817 and 6,391,452, each ofwhich is specifically incorporated by reference. Formulations thatinclude a compound of any of Formula (I), (Ia)-(Ic), (IIa)-(IId),(IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId),(VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh)which are prepared according to these and other techniques well-known inthe art are prepared as solutions in saline, employing benzyl alcohol orother suitable preservatives, fluorocarbons, and/or other solubilizingor dispersing agents known in the art. See, for example, Ansel, H. C. etal., Pharmaceutical Dosage Forms and Drug Delivery Systems, Sixth Ed.(1995). Preferably these compositions and formulations are prepared withsuitable nontoxic pharmaceutically acceptable ingredients. Theseingredients are known to those skilled in the preparation of nasaldosage forms and some of these can be found in REMINGTON: THE SCIENCEAND PRACTICE OF PHARMACY, 21st edition, 2005, a standard reference inthe field. The choice of suitable carriers is highly dependent upon theexact nature of the nasal dosage form desired, e.g., solutions,suspensions, ointments, or gels. Nasal dosage forms generally containlarge amounts of water in addition to the active ingredient. Minoramounts of other ingredients such as pH adjusters, emulsifiers ordispersing agents, preservatives, surfactants, gelling agents, orbuffering and other stabilizing and solubilizing agents may also bepresent. The nasal dosage form should be isotonic with nasal secretions.

For administration by inhalation, the compounds of any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh), described herein may be in a form as anaerosol, a mist or a powder. Pharmaceutical compositions describedherein are conveniently delivered in the form of an aerosol spraypresentation from pressurized packs or a nebuliser, with the use of asuitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol, the dosageunit may be determined by providing a valve to deliver a metered amount.Capsules and cartridges of, such as, by way of example only, gelatin foruse in an inhaler or insufflator may be formulated containing a powdermix of the compound described herein and a suitable powder base such aslactose or starch.

Buccal Formulations

Buccal formulations that include compounds of any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh) may be administered using a variety offormulations known in the art. For example, such formulations include,but are not limited to, U.S. Pat. Nos. 4,229,447, 4,596,795, 4,755,386,and 5,739,136, each of which is specifically incorporated by reference.In addition, the buccal dosage forms described herein can furtherinclude a bioerodible (hydrolysable) polymeric carrier that also servesto adhere the dosage form to the buccal mucosa. The buccal dosage formis fabricated so as to erode gradually over a predetermined time period,wherein the delivery of the compound of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh), is provided essentially throughout. Buccal drug delivery,as will be appreciated by those skilled in the art, avoids thedisadvantages encountered with oral drug administration, e.g., slowabsorption, degradation of the active agent by fluids present in thegastrointestinal tract and/or first-pass inactivation in the liver. Withregard to the bioerodible (hydrolysable) polymeric carrier, it will beappreciated that virtually any such carrier can be used, so long as thedesired drug release profile is not compromised, and the carrier iscompatible with the compound of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh), and any other components that may be present in thebuccal dosage unit. Generally, the polymeric carrier compriseshydrophilic (water-soluble and water-swellable) polymers that adhere tothe wet surface of the buccal mucosa. Examples of polymeric carriersuseful herein include acrylic acid polymers and co, e.g., those known as“carbomers” (Carbopol®, which may be obtained from B.F. Goodrich, is onesuch polymer). Other components may also be incorporated into the buccaldosage forms described herein include, but are not limited to,disintegrants, diluents, binders, lubricants, flavoring, colorants,preservatives, and the like. For buccal or sublingual administration,the compositions may take the form of tablets, lozenges, or gelsformulated in a conventional manner.

Transdermal Formulations

Transdermal formulations described herein may be administered using avariety of devices which have been described in the art. For example,such devices include, but are not limited to, U.S. Pat. Nos. 3,598,122,3,598,123, 3,710,795, 3,731,683, 3,742,951, 3,814,097, 3,921,636,3,972,995, 3,993,072, 3,993,073, 3,996,934, 4,031,894, 4,060,084,4,069,307, 4,077,407, 4,201,211, 4,230,105, 4,292,299, 4,292,303,5,336,168, 5,665,378, 5,837,280, 5,869,090, 6,923,983, 6,929,801 and6,946,144, each of which is specifically incorporated by reference inits entirety.

The transdermal dosage forms described herein may incorporate certainpharmaceutically acceptable excipients which are conventional in theart. In one embodiment, the transdermal formulations described hereininclude at least three components: (1) a formulation of a compound ofany of Formula (I); (2) a penetration enhancer; and (3) an aqueousadjuvant. In addition, transdermal formulations can include additionalcomponents such as, but not limited to, gelling agents, creams andointment bases, and the like. In some embodiments, the transdermalformulation can further include a woven or non-woven backing material toenhance absorption and prevent the removal of the transdermalformulation from the skin. In other embodiments, the transdermalformulations described herein can maintain a saturated or supersaturatedstate to promote diffusion into the skin.

Formulations suitable for transdermal administration of compoundsdescribed herein may employ transdermal delivery devices and transdermaldelivery patches and can be lipophilic emulsions or buffered, aqueoussolutions, dissolved and/or dispersed in a polymer or an adhesive. Suchpatches may be constructed for continuous, pulsatile, or on demanddelivery of pharmaceutical agents. Still further, transdermal deliveryof the compounds described herein can be accomplished by means ofiontophoretic patches and the like. Additionally, transdermal patchescan provide controlled delivery of the compounds of any of Formula (I),(Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd),(VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh),(XIa)-(XIj) or (XIIa)-(XIIh). The rate of absorption can be slowed byusing rate-controlling membranes or by trapping the compound within apolymer matrix or gel. Conversely, absorption enhancers can be used toincrease absorption. An absorption enhancer or carrier can includeabsorbable pharmaceutically acceptable solvents to assist passagethrough the skin. For example, transdermal devices are in the form of abandage comprising a backing member, a reservoir containing the compoundoptionally with carriers, optionally a rate controlling barrier todeliver the compound to the skin of the host at a controlled andpredetermined rate over a prolonged period of time, and means to securethe device to the skin.

Injectable Formulations

Formulations that include a compound of any of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh), suitable for intramuscular, subcutaneous, or intravenousinjection may include physiologically acceptable sterile aqueous ornon-aqueous solutions, dispersions, suspensions or emulsions, andsterile powders for reconstitution into sterile injectable solutions ordispersions. Examples of suitable aqueous and non-aqueous carriers,diluents, solvents, or vehicles including water, ethanol, polyols(propyleneglycol, polyethylene-glycol, glycerol, cremophor and thelike), suitable mixtures thereof, vegetable oils (such as olive oil) andinjectable organic esters such as ethyl oleate. Proper fluidity can bemaintained, for example, by the use of a coating such as lecithin, bythe maintenance of the required particle size in the case ofdispersions, and by the use of surfactants. Formulations suitable forsubcutaneous injection may also contain additives such as preserving,wetting, emulsifying, and dispensing agents. Prevention of the growth ofmicroorganisms can be ensured by various antibacterial and antifungalagents, such as parabens, chlorobutanol, phenol, sorbic acid, and thelike. It may also be desirable to include isotonic agents, such assugars, sodium chloride, and the like. Prolonged absorption of theinjectable pharmaceutical form can be brought about by the use of agentsdelaying absorption, such as aluminum monostearate and gelatin.

For intravenous injections, compounds described herein may be formulatedin aqueous solutions, preferably in physiologically compatible bufferssuch as Hank's solution, Ringer's solution, or physiological salinebuffer. For transmucosal administration, penetrants appropriate to thebarrier to be permeated are used in the formulation. Such penetrants aregenerally known in the art. For other parenteral injections, appropriateformulations may include aqueous or nonaqueous solutions, preferablywith physiologically compatible buffers or excipients. Such excipientsare generally known in the art.

Parenteral injections may involve bolus injection or continuousinfusion. Formulations for injection may be presented in unit dosageform, e.g., in ampoules or in multi-dose containers, with an addedpreservative. The pharmaceutical composition described herein may be ina form suitable for parenteral injection as a sterile suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilizing and/or dispersingagents. Pharmaceutical compositions for parenteral administrationinclude aqueous solutions of the active compounds in water-soluble form.Additionally, suspensions of the active compounds may be prepared asappropriate oily injection suspensions. Suitable lipophilic solvents orvehicles include fatty oils such as sesame oil, or synthetic fatty acidesters, such as ethyl oleate or triglycerides, or liposomes. Aqueousinjection suspensions may contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use.

Other Formulations

In certain embodiments, delivery systems for pharmaceutical compoundsmay be employed, such as, for example, liposomes and emulsions. Incertain embodiments, compositions provided herein can also include anmucoadhesive polymer, selected from among, for example,carboxymethylcellulose, carbomer (acrylic acid polymer),poly(methylmethacrylate), polyacrylamide, polycarbophil, acrylicacid/butyl acrylate copolymer, sodium alginate and dextran.

In some embodiments, the compounds described herein may be administeredtopically and can be formulated into a variety of topicallyadministrable compositions, such as solutions, suspensions, lotions,gels, pastes, medicated sticks, balms, creams or ointments. Suchpharmaceutical compounds can contain solubilizers, stabilizers, tonicityenhancing agents, buffers and preservatives.

The compounds described herein may also be formulated in rectalcompositions such as enemas, rectal gels, rectal foams, rectal aerosols,suppositories, jelly suppositories, or retention enemas, containingconventional suppository bases such as cocoa butter or other glycerides,as well as synthetic polymers such as polyvinylpyrrolidone, PEG, and thelike. In suppository forms of the compositions, a low-melting wax suchas, but not limited to, a mixture of fatty acid glycerides, optionallyin combination with cocoa butter is first melted.

Examples of Methods of Dosing and Treatment Regimens

The compounds described herein can be used in the preparation ofmedicaments for the inhibition of Btk or a homolog thereof, or for thetreatment of diseases or conditions that would benefit, at least inpart, from inhibition of Btk or a homolog thereof. In addition, a methodfor treating any of the diseases or conditions described herein in asubject in need of such treatment, involves administration ofpharmaceutical compositions containing at least one compound of any ofFormula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh), described herein, or apharmaceutically acceptable salt, pharmaceutically acceptable N-oxide,pharmaceutically active metabolite, pharmaceutically acceptable prodrug,or pharmaceutically acceptable solvate thereof, in therapeuticallyeffective amounts to said subject.

The compositions containing the compound(s) described herein can beadministered for prophylactic and/or therapeutic treatments. Intherapeutic applications, the compositions are administered to a patientalready suffering from a disease or condition, in an amount sufficientto cure or at least partially arrest the symptoms of the disease orcondition. Amounts effective for this use will depend on the severityand course of the disease or condition, previous therapy, the patient'shealth status, weight, and response to the drugs, and the judgment ofthe treating physician. It is considered well within the skill of theart for one to determine such therapeutically effective amounts byroutine experimentation (including, but not limited to, a doseescalation clinical trial).

In prophylactic applications, compositions containing the compoundsdescribed herein are administered to a patient susceptible to orotherwise at risk of a particular disease, disorder or condition. Suchan amount is defined to be a “prophylactically effective amount ordose.” In this use, the precise amounts also depend on the patient'sstate of health, weight, and the like. It is considered well within theskill of the art for one to determine such prophylactically effectiveamounts by routine experimentation (e.g., a dose escalation clinicaltrial). When used in a patient, effective amounts for this use willdepend on the severity and course of the disease, disorder or condition,previous therapy, the patient's health status and response to the drugs,and the judgment of the treating physician.

In the case wherein the patient's condition does not improve, upon thedoctor's discretion the administration of the compounds may beadministered chronically, that is, for an extended period of time,including throughout the duration of the patient's life in order toameliorate or otherwise control or limit the symptoms of the patient'sdisease or condition.

In the case wherein the patient's status does improve, upon the doctor'sdiscretion the administration of the compounds may be givencontinuously; alternatively, the dose of drug being administered may betemporarily reduced or temporarily suspended for a certain length oftime (i.e., a “drug holiday”). The length of the drug holiday can varybetween 2 days and 1 year, including by way of example only, 2 days, 3days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days,180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or365 days. The dose reduction during a drug holiday may be from 10%-100%,including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%,45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, the dosage or thefrequency of administration, or both, can be reduced, as a function ofthe symptoms, to a level at which the improved disease, disorder orcondition is retained. Patients can, however, require intermittenttreatment on a long-term basis upon any recurrence of symptoms.

The amount of a given agent that will correspond to such an amount willvary depending upon factors such as the particular compound, disease orcondition and its severity, the identity (e.g., weight) of the subjector host in need of treatment, but can nevertheless be routinelydetermined in a manner known in the art according to the particularcircumstances surrounding the case, including, e.g., the specific agentbeing administered, the route of administration, the condition beingtreated, and the subject or host being treated. In general, however,doses employed for adult human treatment will typically be in the rangeof 0.02-5000 mg per day, or from about 1-1500 mg per day. The desireddose may conveniently be presented in a single dose or as divided dosesadministered simultaneously (or over a short period of time) or atappropriate intervals, for example as two, three, four or more sub-dosesper day.

The pharmaceutical composition described herein may be in unit dosageforms suitable for single administration of precise dosages. In unitdosage form, the formulation is divided into unit doses containingappropriate quantities of one or more compound. The unit dosage may bein the form of a package containing discrete quantities of theformulation. Non-limiting examples are packaged tablets or capsules, andpowders in vials or ampoules. Aqueous suspension compositions can bepackaged in single-dose non-reclosable containers. Alternatively,multiple-dose reclosable containers can be used, in which case it istypical to include a preservative in the composition. By way of exampleonly, formulations for parenteral injection may be presented in unitdosage form, which include, but are not limited to ampoules, or inmulti-dose containers, with an added preservative.

The foregoing ranges are merely suggestive, as the number of variablesin regard to an individual treatment regime is large, and considerableexcursions from these recommended values are not uncommon Such dosagesmay be altered depending on a number of variables, not limited to theactivity of the compound used, the disease or condition to be treated,the mode of administration, the requirements of the individual subject,the severity of the disease or condition being treated, and the judgmentof the practitioner.

Toxicity and therapeutic efficacy of such therapeutic regimens can bedetermined by standard pharmaceutical procedures in cell cultures orexperimental animals, including, but not limited to, the determinationof the LD₅₀ (the dose lethal to 50% of the population) and the ED₅₀ (thedose therapeutically effective in 50% of the population). The dose ratiobetween the toxic and therapeutic effects is the therapeutic index andit can be expressed as the ratio between LD₅₀ and ED₅₀. Compoundsexhibiting high therapeutic indices are preferred. The data obtainedfrom cell culture assays and animal studies can be used in formulating arange of dosage for use in human. The dosage of such compounds liespreferably within a range of circulating concentrations that include theED₅₀ with minimal toxicity. The dosage may vary within this rangedepending upon the dosage form employed and the route of administrationutilized.

Combination Treatments

The irreversible Btk inhibitor compositions described herein can also beused in combination with other well known therapeutic reagents that areselected for their therapeutic value for the condition to be treated. Ingeneral, the compositions described herein and, in embodiments wherecombinational therapy is employed, other agents do not have to beadministered in the same pharmaceutical composition, and may, because ofdifferent physical and chemical characteristics, have to be administeredby different routes. The determination of the mode of administration andthe advisability of administration, where possible, in the samepharmaceutical composition, is well within the knowledge of the skilledclinician. The initial administration can be made according toestablished protocols known in the art, and then, based upon theobserved effects, the dosage, modes of administration and times ofadministration can be modified by the skilled clinician.

In certain instances, it may be appropriate to administer at least oneirreversible Btk inhibitor compound described herein in combination withanother therapeutic agent. By way of example only, if one of the sideeffects experienced by a patient upon receiving one of the irreversibleBtk inhibitor compounds described herein is nausea, then it may beappropriate to administer an anti-nausea agent in combination with theinitial therapeutic agent. Or, by way of example only, the therapeuticeffectiveness of one of the compounds described herein may be enhancedby administration of an adjuvant (i.e., by itself the adjuvant may haveminimal therapeutic benefit, but in combination with another therapeuticagent, the overall therapeutic benefit to the patient is enhanced). Or,by way of example only, the benefit experienced by a patient may beincreased by administering one of the compounds described herein withanother therapeutic agent (which also includes a therapeutic regimen)that also has therapeutic benefit. In any case, regardless of thedisease, disorder or condition being treated, the overall benefitexperienced by the patient may simply be additive of the two therapeuticagents or the patient may experience a synergistic benefit.

The particular choice of compounds used will depend upon the diagnosisof the attending physicians and their judgment of the condition of thepatient and the appropriate treatment protocol. The compounds may beadministered concurrently (e.g., simultaneously, essentiallysimultaneously or within the same treatment protocol) or sequentially,depending upon the nature of the disease, disorder, or condition, thecondition of the patient, and the actual choice of compounds used. Thedetermination of the order of administration, and the number ofrepetitions of administration of each therapeutic agent during atreatment protocol, is well within the knowledge of the skilledphysician after evaluation of the disease being treated and thecondition of the patient.

It is known to those of skill in the art that therapeutically-effectivedosages can vary when the drugs are used in treatment combinations.Methods for experimentally determining therapeutically-effective dosagesof drugs and other agents for use in combination treatment regimens aredescribed in the literature. For example, the use of metronomic dosing,i.e., providing more frequent, lower doses in order to minimize toxicside effects, has been described extensively in the literatureCombination treatment further includes periodic treatments that startand stop at various times to assist with the clinical management of thepatient.

For combination therapies described herein, dosages of theco-administered compounds will of course vary depending on the type ofco-drug employed, on the specific drug employed, on the disease orcondition being treated and so forth. In addition, when co-administeredwith one or more biologically active agents, the compound providedherein may be administered either simultaneously with the biologicallyactive agent(s), or sequentially. If administered sequentially, theattending physician will decide on the appropriate sequence ofadministering protein in combination with the biologically activeagent(s).

In any case, the multiple therapeutic agents (one of which is a compoundof Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh), described herein) may beadministered in any order or even simultaneously. If simultaneously, themultiple therapeutic agents may be provided in a single, unified form,or in multiple forms (by way of example only, either as a single pill oras two separate pills). One of the therapeutic agents may be given inmultiple doses, or both may be given as multiple doses. If notsimultaneous, the timing between the multiple doses may vary from morethan zero weeks to less than four weeks. In addition, the combinationmethods, compositions and formulations are not to be limited to the useof only two agents; the use of multiple therapeutic combinations arealso envisioned.

It is understood that the dosage regimen to treat, prevent, orameliorate the condition(s) for which relief is sought, can be modifiedin accordance with a variety of factors. These factors include thedisorder from which the subject suffers, as well as the age, weight,sex, diet, and medical condition of the subject. Thus, the dosageregimen actually employed can vary widely and therefore can deviate fromthe dosage regimens set forth herein.

The pharmaceutical agents which make up the combination therapydisclosed herein may be a combined dosage form or in separate dosageforms intended for substantially simultaneous administration. Thepharmaceutical agents that make up the combination therapy may also beadministered sequentially, with either therapeutic compound beingadministered by a regimen calling for two-step administration. Thetwo-step administration regimen may call for sequential administrationof the active agents or spaced-apart administration of the separateactive agents. The time period between the multiple administration stepsmay range from, a few minutes to several hours, depending upon theproperties of each pharmaceutical agent, such as potency, solubility,bioavailability, plasma half-life and kinetic profile of thepharmaceutical agent. Circadian variation of the target moleculeconcentration may also determine the optimal dose interval.

In addition, the compounds described herein also may be used incombination with procedures that may provide additional or synergisticbenefit to the patient. By way of example only, patients are expected tofind therapeutic and/or prophylactic benefit in the methods describedherein, wherein pharmaceutical composition of a compound disclosedherein and/or combinations with other therapeutics are combined withgenetic testing to determine whether that individual is a carrier of amutant gene that is known to be correlated with certain diseases orconditions.

The compounds described herein and combination therapies can beadministered before, during or after the occurrence of a disease orcondition, and the timing of administering the composition containing acompound can vary. Thus, for example, the compounds can be used as aprophylactic and can be administered continuously to subjects with apropensity to develop conditions or diseases in order to prevent theoccurrence of the disease or condition. The compounds and compositionscan be administered to a subject during or as soon as possible after theonset of the symptoms. The administration of the compounds can beinitiated within the first 48 hours of the onset of the symptoms, withinthe first 6 hours of the onset of the symptoms, or within 3 hours of theonset of the symptoms. The initial administration can be via any routepractical, such as, for example, an intravenous injection, a bolusinjection, infusion over 5 minutes to about 5 hours, a pill, a capsule,transdermal patch, buccal delivery, and the like, or combinationthereof. A compound should be administered as soon as is practicableafter the onset of a disease or condition is detected or suspected, andfor a length of time necessary for the treatment of the disease, suchas, for example, from about 1 month to about 3 months. The length oftreatment can vary for each subject, and the length can be determinedusing the known criteria. For example, the compound or a formulationcontaining the compound can be administered for at least 2 weeks,between about 1 month to about 5 years, or from about 1 month to about 3years.

Exemplary Therapeutic Agents for Use in Combination with an IrreversibleBtk Inhibitor Compound

Where the subject is suffering from or at risk of suffering from anautoimmune disease, an inflammatory disease, or an allergy disease, anirreversible Btk inhibitor compound can be used in with one or more ofthe following therapeutic agents in any combination: immunosuppressants(e.g., tacrolimus, cyclosporin, rapamicin, methotrexate,cyclophosphamide, azathioprine, mercaptopurine, mycophenolate, orFTY720), glucocorticoids (e.g., prednisone, cortisone acetate,prednisolone, methylprednisolone, dexamethasone, betamethasone,triamcinolone, beclometasone, fludrocortisone acetate,deoxycorticosterone acetate, aldosterone), non-steroidalanti-inflammatory drugs (e.g., salicylates, arylalkanoic acids,2-arylpropionic acids, N-arylanthranilic acids, oxicams, coxibs, orsulphonanilides), Cox-2-specific inhibitors (e.g., valdecoxib,celecoxib, or rofecoxib), leflunomide, gold thioglucose, goldthiomalate, aurofin, sulfasalazine, hydroxychloroquinine, minocycline,TNF-α binding proteins (e.g., infliximab, etanercept, or adalimumab),abatacept, anakinra, interferon-β, interferon-γ, interleukin-2, allergyvaccines, antihistamines, antileukotrienes, beta-agonists, theophylline,or anticholinergics.

Where the subject is suffering from or at risk of suffering from aB-cell proliferative disorder (e.g., plasma cell myeloma), the subjectedcan be treated with an irreversible Btk inhibitor compound in anycombination with one or more other anti-cancer agents. In someembodiments, one or more of the anti-cancer agents are proapoptoticagents. Examples of anti-cancer agents include, but are not limited to,any of the following: gossyphol, genasense, polyphenol E, Chlorofusin,all trans-retinoic acid (ATRA), bryostatin, tumor necrosisfactor-related apoptosis-inducing ligand (TRAIL),5-aza-2′-deoxycytidine, all trans retinoic acid, doxorubicin,vincristine, etoposide, gemcitabine, imatinib (Gleevec®), geldanamycin,17-N-Allylamino-17-Demethoxygeldanamycin (17-AAG), flavopiridol,LY294002, bortezomib, trastuzumab, BAY 11-7082, PKC412, or PD184352,Taxol™, also referred to as “paclitaxel”, which is a well-knownanti-cancer drug which acts by enhancing and stabilizing microtubuleformation, and analogs of Taxol™, such as Taxotere™. Compounds that havethe basic taxane skeleton as a common structure feature, have also beenshown to have the ability to arrest cells in the G2-M phases due tostabilized microtubules and may be useful for treating cancer incombination with the compounds described herein.

Further examples of anti-cancer agents for use in combination with anirreversible Btk inhibitor compound include inhibitors ofmitogen-activated protein kinase signaling, e.g., U0126, PD98059,PD184352, PD0325901, ARRY-142886, SB239063, SP600125, BAY 43-9006,wortmannin, or LY294002; Syk inhibitors; mTOR inhibitors; and antibodies(e.g., rituxan).

Other anti-cancer agents that can be employed in combination with anirreversible Btk inhibitor compound include Adriamycin, Dactinomycin,Bleomycin, Vinblastine, Cisplatin, acivicin; aclarubicin; acodazolehydrochloride; acronine; adozelesin; aldesleukin; altretamine;ambomycin; ametantrone acetate; aminoglutethimide; amsacrine;anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa;azotomycin; batimastat; benzodepa; bicalutamide; bisantrenehydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate;brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone;caracemide; carbetimer; carboplatin; carmustine; carubicinhydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin;cladribine; crisnatol mesylate; cyclophosphamide; cytarabine;dacarbazine; daunorubicin hydrochloride; decitabine; dexormaplatin;dezaguanine; dezaguanine mesylate; diaziquone; doxorubicin; doxorubicinhydrochloride; droloxifene; droloxifene citrate; dromostanolonepropionate; duazomycin; edatrexate; eflornithine hydrochloride;elsamitrucin; enloplatin; enpromate; epipropidine; epirubicinhydrochloride; erbulozole; esorubicin hydrochloride; estramustine;estramustine phosphate sodium; etanidazole; etoposide; etoposidephosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide;floxuridine; fludarabine phosphate; fluorouracil; flurocitabine;fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride;hydroxyurea; idarubicin hydrochloride; ifosfamide; iimofosine;interleukin Il (including recombinant interleukin II, or rlL2),interferon α-2a; interferon α-2b; interferon α-n1; interferon α-n3;interferon β-la; interferon γ-lb; iproplatin; irinotecan hydrochloride;lanreotide acetate; letrozole; leuprolide acetate; liarozolehydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride;masoprocol; maytansine; mechlorethamine hydrochloride; megestrolacetate; melengestrol acetate; melphalan; menogaril; mercaptopurine;methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide;mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper;mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazoie;nogalamycin; ormaplatin; oxisuran; pegaspargase; peliomycin;pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan;piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium;porfiromycin; prednimustine; procarbazine hydrochloride; puromycin;puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol;safingol hydrochloride; semustine; simtrazene; sparfosate sodium;sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin;streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium;tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicinhydrochloride.

Other anti-cancer agents that can be employed in combination with anirreversible Btk inhibitor compound include: 20-epi-1, 25dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin;acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists;altretamine; ambamustine; amidox; amifostine; aminolevulinic acid;amrubicin; amsacrine; anagrelide; anastrozole; andrographolide;angiogenesis inhibitors; antagonist D; antagonist G; antarelix;anti-dorsalizing morphogenetic protein-1; antiandrogen, prostaticcarcinoma; antiestrogen; antineoplaston; antisense oligonucleotides;aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators;apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine;atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3;azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol;batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine;beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid;bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine;bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane;buthionine sulfoximine; calcipotriol; calphostin C; camptothecinderivatives; canarypox IL-2; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;9-dioxamycin; diphenyl spiromustine; docosanol; dolasetron;doxifluridine; droloxifene; dronabinol; duocarmycin SA; ebselen;ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur;epirubicin; epristeride; estramustine analogue; estrogen agonists;estrogen antagonists; etanidazole; etoposide phosphate; exemestane;fadrozole; fazarabine; fenretinide; filgrastim; finasteride;flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;imidazoacridones; imiquimod; immunostimulant peptides; insulin-likegrowth factor-1 receptor inhibitor; interferon agonists; interferons;interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact;irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin;pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine;pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin;pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylerie conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen-binding protein; sizofiran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatinstimalamer.

Yet other anticancer agents that can be employed in combination with anirreversible Btk inhibitor compound include alkylating agents,antimetabolites, natural products, or hormones, e.g., nitrogen mustards(e.g., mechloroethamine, cyclophosphamide, chlorambucil, etc.), alkylsulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine, lomusitne,etc.), or triazenes (decarbazine, etc.). Examples of antimetabolitesinclude but are not limited to folic acid analog (e.g., methotrexate),or pyrimidine analogs (e.g., Cytarabine), purine analogs (e.g.,mercaptopurine, thioguanine, pentostatin).

Examples of natural products useful in combination with an irreversibleBtk inhibitor compound include but are not limited to vinca alkaloids(e.g., vinblastin, vincristine), epipodophyllotoxins (e.g., etoposide),antibiotics (e.g., daunorubicin, doxorubicin, bleomycin), enzymes (e.g.,L-asparaginase), or biological response modifiers (e.g., interferonalpha).

Examples of alkylating agents that can be employed in combination anirreversible Btk inhibitor compound include, but are not limited to,nitrogen mustards (e.g., mechloroethamine, cyclophosphamide,chlorambucil, meiphalan, etc.), ethylenimine and methylmelamines (e.g.,hexamethlymelamine, thiotepa), alkyl sulfonates (e.g., busulfan),nitrosoureas (e.g., carmustine, lomusitne, semustine, streptozocin,etc.), or triazenes (decarbazine, etc.). Examples of antimetabolitesinclude, but are not limited to folic acid analog (e.g., methotrexate),or pyrimidine analogs (e.g., fluorouracil, floxouridine, Cytarabine),purine analogs (e.g., mercaptopurine, thioguanine, pentostatin.

Examples of hormones and antagonists useful in combination with anirreversible Btk inhibitor compound include, but are not limited to,adrenocorticosteroids (e.g., prednisone), progestins (e.g.,hydroxyprogesterone caproate, megestrol acetate, medroxyprogesteroneacetate), estrogens (e.g., diethlystilbestrol, ethinyl estradiol),antiestrogen (e.g., tamoxifen), androgens (e.g., testosteronepropionate, fluoxymesterone), antiandrogen (e.g., flutamide),gonadotropin releasing hormone analog (e.g., leuprolide). Other agentsthat can be used in the methods and compositions described herein forthe treatment or prevention of cancer include platinum coordinationcomplexes (e.g., cisplatin, carboblatin), anthracenedione (e.g.,mitoxantrone), substituted urea (e.g., hydroxyurea), methyl hydrazinederivative (e.g., procarbazine), adrenocortical suppressant (e.g.,mitotane, aminoglutethimide).

Examples of anti-cancer agents which act by arresting cells in the G2-Mphases due to stabilized microtubules and which can be used incombination with an irreversible Btk inhibitor compound include withoutlimitation the following marketed drugs and drugs in development:Erbulozole (also known as R-55104), Dolastatin 10 (also known as DLS-10and NSC-376128), Mivobulin isethionate (also known as CI-980),Vincristine, NSC-639829, Discodermolide (also known as NVP-XX-A-296),ABT-751 (Abbott, also known as E-7010), Altorhyrtins (such asAltorhyrtin A and Altorhyrtin C), Spongistatins (such as Spongistatin 1,Spongistatin 2, Spongistatin 3, Spongistatin 4, Spongistatin 5,Spongistatin 6, Spongistatin 7, Spongistatin 8, and Spongistatin 9),Cemadotin hydrochloride (also known as LU-103793 and NSC-D-669356),Epothilones (such as Epothilone A, Epothilone B, Epothilone C (alsoknown as desoxyepothilone A or dEpoA), Epothilone D (also referred to asKOS-862, dEpoB, and desoxyepothilone B), Epothilone E, Epothilone F,Epothilone B N-oxide, Epothilone A N-oxide, 16-aza-epothilone B,21-aminoepothilone B (also known as BMS-310705), 21-hydroxyepothilone D(also known as Desoxyepothilone F and dEpoF), 26-fluoroepothilone),Auristatin PE (also known as NSC-654663), Soblidotin (also known asTZT-1027), LS-4559-P (Pharmacia, also known as LS-4577), LS-4578(Pharmacia, also known as LS-477-P), LS-4477 (Pharmacia), LS-4559(Pharmacia), RPR-112378 (Aventis), Vincristine sulfate, DZ-3358(Daiichi), FR-182877 (Fujisawa, also known as WS-9885B), GS-164(Takeda), GS-198 (Takeda), KAR-2 (Hungarian Academy of Sciences),BSF-223651 (BASF, also known as ILX-651 and LU-223651), SAH-49960(Lilly/Novartis), SDZ-268970 (Lilly/Novartis), AM-97 (Armad/KyowaHakko), AM-132 (Armad), AM-138 (Armad/Kyowa Hakko), IDN-5005 (Indena),Cryptophycin 52 (also known as LY-355703), AC-7739 (Ajinomoto, alsoknown as AVE-8063A and CS-39.HCI), AC-7700 (Ajinomoto, also known asAVE-8062, AVE-8062A, CS-39-L-Ser.HCI, and RPR-258062A), Vitilevuamide,Tubulysin A, Canadensol, Centaureidin (also known as NSC-106969),T-138067 (Tularik, also known as T-67, TL-138067 and TI-138067), COBRA-1(Parker Hughes Institute, also known as DDE-261 and WHI-261), H10(Kansas State University), H16 (Kansas State University), Oncocidin A1(also known as BTO-956 and DIME), DDE-313 (Parker Hughes Institute),Fijianolide B, Laulimalide, SPA-2 (Parker Hughes Institute), SPA-1(Parker Hughes Institute, also known as SPIKET-P), 3-IAABU(Cytoskeleton/Mt. Sinai School of Medicine, also known as MF-569),Narcosine (also known as NSC-5366), Nascapine, D-24851 (Asta Medica),A-105972 (Abbott), Hemiasterlin, 3-BAABU (Cytoskeleton/Mt. Sinai Schoolof Medicine, also known as MF-191), TMPN (Arizona State University),Vanadocene acetylacetonate, T-138026 (Tularik), Monsatrol, lnanocine(also known as NSC-698666), 3-1AABE (Cytoskeleton/Mt. Sinai School ofMedicine), A-204197 (Abbott), T-607 (Tuiarik, also known as T-900607),RPR-115781 (Aventis), Eleutherobins (such as Desmethyleleutherobin,Desaetyleleutherobin, lsoeleutherobin A, and Z-Eleutherobin),Caribaeoside, Caribaeolin, Halichondrin B, D-64131 (Asta Medica),D-68144 (Asta Medica), Diazonamide A, A-293620 (Abbott), NPI-2350(Nereus), Taccalonolide A, TUB-245 (Aventis), A-259754 (Abbott),Diozostatin, (−)-Phenylahistin (also known as NSCL-96F037), D-68838(Asta Medica), D-68836 (Asta Medica), Myoseverin B, D-43411 (Zentaris,also known as D-81862), A-289099 (Abbott), A-318315 (Abbott), HTI-286(also known as SPA-110, trifluoroacetate salt) (Wyeth), D-82317(Zentaris), D-82318 (Zentaris), SC-12983 (NCI), Resverastatin phosphatesodium, BPR-OY-007 (National Health Research Institutes), and SSR-250411(Sanofi).

Where the subject is suffering from or at risk of suffering from athromboembolic disorder (e.g., stroke), the subject can be treated withan irreversible Btk inhibitor compound in any combination with one ormore other anti-thromboembolic agents. Examples of anti-thromboembolicagents include, but are not limited any of the following: thrombolyticagents (e.g., alteplase anistreplase, streptokinase, urokinase, ortissue plasminogen activator), heparin, tinzaparin, warfarin, dabigatran(e.g., dabigatran etexilate), factor Xa inhibitors (e.g., fondaparinux,draparinux, rivaroxaban, DX-9065a, otamixaban, LY517717, or YM150),ticlopidine, clopidogrel, CS-747 (prasugrel, LY640315), ximelagatran, orBIBR 1048.

Kits/Articles of Manufacture

For use in the therapeutic applications described herein, kits andarticles of manufacture are also described herein. Such kits can includea carrier, package, or container that is compartmentalized to receiveone or more containers such as vials, tubes, and the like, each of thecontainer(s) including one of the separate elements to be used in amethod described herein. Suitable containers include, for example,bottles, vials, syringes, and test tubes. The containers can be formedfrom a variety of materials such as glass or plastic.

The articles of manufacture provided herein contain packaging materials.Packaging materials for use in packaging pharmaceutical products arewell known to those of skill in the art. See, e.g., U.S. Pat. Nos.5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical packagingmaterials include, but are not limited to, blister packs, bottles,tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, andany packaging material suitable for a selected formulation and intendedmode of administration and treatment. A wide array of formulations ofthe compounds and compositions provided herein are contemplated as are avariety of treatments for any disease, disorder, or condition that wouldbenefit by inhibition of Btk, or in which Btk is a mediator orcontributor to the symptoms or cause.

For example, the container(s) can include one or more compoundsdescribed herein, optionally in a composition or in combination withanother agent as disclosed herein. The container(s) optionally have asterile access port (for example the container can be an intravenoussolution bag or a vial having a stopper pierceable by a hypodermicinjection needle). Such kits optionally comprising a compound with anidentifying description or label or instructions relating to its use inthe methods described herein.

A kit will typically may include one or more additional containers, eachwith one or more of various materials (such as reagents, optionally inconcentrated form, and/or devices) desirable from a commercial and userstandpoint for use of a compound described herein. Non-limiting examplesof such materials include, but not limited to, buffers, diluents,filters, needles, syringes; carrier, package, container, vial and/ortube labels listing contents and/or instructions for use, and packageinserts with instructions for use. A set of instructions will alsotypically be included.

A label can be on or associated with the container. A label can be on acontainer when letters, numbers or other characters forming the labelare attached, molded or etched into the container itself; a label can beassociated with a container when it is present within a receptacle orcarrier that also holds the container, e.g., as a package insert. Alabel can be used to indicate that the contents are to be used for aspecific therapeutic application. The label can also indicate directionsfor use of the contents, such as in the methods described herein.

In certain embodiments, the pharmaceutical compositions can be presentedin a pack or dispenser device which can contain one or more unit dosageforms containing a compound provided herein. The pack can for examplecontain metal or plastic foil, such as a blister pack. The pack ordispenser device can be accompanied by instructions for administration.The pack or dispenser can also be accompanied with a notice associatedwith the container in form prescribed by a governmental agencyregulating the manufacture, use, or sale of pharmaceuticals, whichnotice is reflective of approval by the agency of the form of the drugfor human or veterinary administration. Such notice, for example, can bethe labeling approved by the U.S. Food and Drug Administration forprescription drugs, or the approved product insert. Compositionscontaining a compound provided herein formulated in a compatiblepharmaceutical carrier can also be prepared, placed in an appropriatecontainer, and labeled for treatment of an indicated condition.

C481S Mutation

The B cell lymphoma is characterized by a plurality of cells having amutant BTK polypeptide. The mutant BTK polypeptides contain one or moreamino acid substitutions that confers resistance to inhibition by acovalent and/or irreversible BTK inhibitor. The modification can be asubstitution or a deletion of the amino acid at amino acid position 481compared to a wild type BTK. Specifically, the modification can be asubstitution of cysteine to serine at amino acid position 481 of the BTKpolypeptide (“C481 S”) as described by Woyach, et al. (Resistancemechanisms for the Bruton's tyrosine kinase inhibitor ibrutinib, N EnglJ Med. 2014, 12; 370(24):2286-94).

EXAMPLES

The following specific and non-limiting examples are to be construed asmerely illustrative, and do not limit the present disclosure in any waywhatsoever. Without further elaboration, it is believed that one skilledin the art can, based on the description herein, utilize the presentdisclosure to its fullest extent. All publications cited herein arehereby incorporated by reference in their entirety. Where reference ismade to a URL or other such identifier or address, it is understood thatsuch identifiers can change and particular information on the internetcan come and go, but equivalent information can be found by searchingthe internet. Reference thereto evidences the availability and publicdissemination of such information.

The examples below as well as throughout the application, the followingabbreviations have the following meanings. If not defined, the termshave their generally accepted meanings.

-   -   aq=aqueous    -   Boc=tert-butyloxycarbonyl    -   t-BuOH=tertiary butanol    -   DCE=1,2-dichloroethane    -   DCM=dichloromethane    -   DIAD=diisopropyl azodicarboxylate    -   DIEA or DIPEA=N,N-diisopropylethylamine    -   DMAP=dimethylaminopyridine    -   DMF=dimethylformamide    -   DMSO=dimethylsulfoxide    -   ESI=electron spray ionization    -   EA=ethyl acetate    -   g=gram    -   HCl=hydrogen chloride    -   HPLC=high performance liquid chromatography    -   hr=hour    -   ¹H NMR=proton nuclear magnetic resonance    -   IPA=isopropyl alcohol    -   KOAc=potassium acetate    -   LC-MS=liquid chromatography mass spectroscopy    -   M=molar    -   MeCN=acetonitrile    -   MeOH=methanol    -   mg=milligram    -   min=minute    -   ml=milliliter    -   mM=millimolar    -   mmol=millimole    -   m.p.=melting point    -   MS=mass spectrometry    -   m/z=mass-to-charge ratio    -   N=normal    -   NIS=N-iodosuccinimide    -   nM=nanomolar    -   nm=nanometer    -   Pd(dppf)Cl₂=[1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)    -   PE=petroleum ether    -   PyBOP=benzotriazol-1-yl-oxytripyrrolidinophosphonium        hexafluorophosphate    -   quant.=quantitative    -   RP=reverse phase    -   rt or r.t.=room temperature    -   Sat.=saturated    -   TEA=triethylamine    -   TFA=trifluoroacetic acid    -   μL=microliter    -   μM=micromolar

Preparation of Intermediates Intermediate 1: Preparation of3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (658-2)

To a solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (658-1)(5.02 g, 37.2 mmol) in DMF (300 ml) was added NIS (12.57 g, 55.9 mol).The resulting mixture was stirred at 80° C. overnight. After cooled downto r.t., the reaction solution was concentrated down to around 150 mland Na₂SO₃ aqueous (sat., 600 ml) was introduced. The precipitate wascollected by filtration, washed with water (500 ml) and dried under highvacuum to provide 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (658-2)(800 g, 82%) as a light yellow solid. LC-MS (ESI): m/z (M+1) 262.

Intermediate 2: Preparation of (R)-tert-butyl3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylate(658-3)

At −10° C. and under N₂ atmosphere, to a solution of3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (658-2) (18.78 g, 71.94mmol), (S)-tert-butyl 3-hydroxypyrrolidine-1-carboxylate (17.51 g, 93.53mmol) and PPh₃ (28.31 g, 107.9 mmol) in anhydrous THF (300 ml) was addedDIAD (21.82 g, 107.92 mmol) dropwise. The reaction mixture wasmaintained below −5° C. during the addition. After addition of DIAD, thereaction mixture was stirred at r.t. for 10 hours followed by beingpartitioned with water (300 ml) and EA (300 ml). The organic layer wasseparated, and the aqueous layer was extracted with EA (200 ml×3). Thecombined organic layer was washed with brine, dried over Na₂SO₄,concentrated and purified by silica gel column chromatography to givethe product-(R)-tert-butyl3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylate(658-3) (15.02 g, 48%) as a yellow solid. LC-MS (ESI): m/z (M+1) 431.1.

Intermediate 4: Preparation of4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide

Intermediate 5: Preparation of4-bromo-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide

To a solution of 4-bromobenzoic acid (21.0 g. 104.47 mmol) and2-amino-4-trifluoromethyl pyridine (25.0 g, 154.22 mmol) in pyridine(300 ml) was added POCl₃ dropwise at rt under cooling by tap water. Thereaction mixture was stirred at rt for another 30 min and poured inice-water (200 ml). The precipitate was collected by filtration, washedwith water and dried under vacuum under high vacuum for 16 hrs to givedesired product-4-bromo-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide.LC-MS (ESI): m/z (M+1) 347.1.

Intermediate 6: Preparation of4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide

A suspension of 4-bromo-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide(13.8 g, 40 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (15.24 g, 60mmol), Pd(dppf)Cl₂ (3.26 g, 40 mmol) and KOAc (7.85 g, 80 mmol) inanhydrous dioxane (200 ml) was degassed with nitrogen for 5 min. Thenthe reaction mixture was heated to 95° C. in a sealed tube for 10 hrs,and filtered with celite. The filtrate was partitioned with EA (200 ml)and water (200 ml). After separation, the aqueous layer was extractedwith EA 3 times. The combined organic layer was dried with MgSO₄,concentrated and purified by silica gel column chromatography to givetheproduct-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide.LC-MS (ESI): m/z (M+1) 393.0.

Intermediate 7: Preparation of (R)-tert-butyl3-(4-amino-3-(4-(4-(trifluoromethyl)pyridin-2-ylcarbamoyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylate(658-4)

Under N₂ atmosphere, to a suspension of (R)-tert-butyl3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylate(658-3) (6.0 g 13.95 mmol),4-(4,4,5,5-tetramethyl-1,3,2-R-2-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide(7.12 g, 18.13 mmol), Cs₂CO₃ (13.63 g, 41.84 mmol) in dioxane/H₂O (50ml/12.5 ml), was added Pd(dppf)Cl₂ (1.139 g, 1.39 mmol). The resultingmixture was stirred at 100° C. for 13 hrs under N₂ atmosphere in sealedtube. After cooled down to r.t., the mixture was filtered through a padof Celite. The filtrate was concentrated to give a residue which waspartitioned with DCM (200 ml) and water (200 ml). The layers wereseparated, and the organic layer was washed with brine, dried overNa₂SO₄ and concentrated in vacuum to give crude product which waspurified by column chromatography (silica gel, 16% to 75% EA in DCM) toprovide (R)-tert-butyl3-(4-amino-3-(4-(4-(trifluoromethyl)pyridin-2-ylcarbamoyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylate(658-4) (3.1 g, 54%) as yellow solid. LC-MS (ESI): m/z (M+1) 569.3.

Intermediate 8: Preparation of(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamidehydrochloride (658-5)

To a solution of (R)-tert-Butyl3-(4-amino-3-(4-(4-(trifluoromethyl)pyridin-2-ylcarbamoyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylate(658-4) (3.1 g, 5.45 mmol) in methanol 20 ml was added HCl/dioxane (4 N,6 ml) at room temperature. The reaction mixture was heated to 40° C. andstirred for 4 hrs, followed by evaporation of solvent to give(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamidehydrochloride (658-5) (quant.) as a white solid. LC-MS (ESI): m/z (M+1)469.2.

Using the methods given above, the following intermediates (Intermediate9-Intermediate 23) were also prepared.

Intermediate 9:(R)-4-(4-amino-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamidehydrochloride

LC-MS (ESI): m/z (M+1) 457.05; M⁺−1=455.25

Intermediate 10:(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamidehydrochloride

LC-MS (ESI): m/z (M+1) 406.95; M⁺−1=405.1

Intermediate 11:(R)-4-(4-amino-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide

LC-MS (ESI): m/z (M+1) 435.05; M⁺−1=433.15

Intermediate 12:(R)-4-(4-amino-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide

LC-MS (ESI): m/z (M+1) 421.05; M⁺−1=419.1

Intermediate 13:(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamidehydrochloride

LC-MS (ESI): m/z (M+1) 443.1; M⁺−1=411.25

Intermediate 14:(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide2,2,2-trifluoroacetate

LC-MS (ESI): m/z (M+1) 426.0; M⁺−1=424.2

Intermediate 15:(R)-4-(4-amino-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide2,2,2-trifluoroacetate

LC-MS (ESI): m/z (M+1) 440.10; M⁺−1=438.2

Intermediate 16:(R)-4-(4-amino-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamidehydrochloride

LC-MS (ESI): m/z (M+1) 483.05; M⁺−1=481.2

Intermediate 17:(R)-4-(4-amino-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamidehydrochloride

LC-MS (ESI): m/z (M+1) 429.05; M⁺−1=427.2

Intermediate 18:(R)-4-(4-amino-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamidehydrochloride

LC-MS (ESI): m/z (M+1) 415.05; M⁺−1=413.20

Intermediate 19:(R)-4-(4-amino-1-(piperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamidehydrochloride

LC-MS (ESI): m/z (M+1) 457.10; M⁺−1=455.25

Intermediate 20:(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamidehydrochloride

LC-MS (ESI): m/z (M+1) 443.10; M⁺−1=441.3

Intermediate 21:(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamidehydrochloride

LC-MS (ESI): m/z (M+1) 401.05

Intermediate 22:(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamidehydrochloride

LC-MS (ESI): m/z (M+1)=421.0; M⁺−1=419.1

Intermediate 23:(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamidehydrochloride

Synthesis of Compounds of the Invention Example 1 Synthesis of(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide(658) (Compound ID 1)

To a solution of (R)-tert-butyl 3-(4-amino-3-(4-(4-(trifluoromethyl)pyridin-2-ylcarbamoyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)pyrrolidine-1-carboxylate(658-5) (57.8 mg, 0.1 mmol), but-2-ynoic acid (12.6 mg, 0.15 mmol) andPyBOP (104 mg, 0.2 mmol) in DMF (1.5 ml) was added DIPEA (77.54 mg, 0.6mmol) dropwise. The reaction mixture was stirred at rt for 30 min.Quenched with NaHCO₃, The reaction mixture was extracted with EA 3times. The resulting organic layer was washed brine and dried overNa₂SO₄. The solvent was removed in vacuum and the residue was purifiedby column chromatography (silica gel, 1% to 3% MeOH in EA) to give 40 mgof(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide,yield 75%. LC-MS (ESI): m/z (M−1) 535.3; ¹H NMR (400 MHz, DMSO) δ 11.38(s, 1H), 8.71 (d, J=5.1 Hz, 1H), 8.58 (s, 1H), 8.31 (d, J=4.0 Hz, 1H),8.23 (d, J=8.3 Hz, 2H), 7.82 (d, J=8.3 Hz, 2H), 7.57 (d, J=5.1 Hz, 1H),5.55 (s, 1H), 4.15-3.53 (m, 4H), 2.46 (dd, J=13.8, 7.2 Hz, 2H), 2.01 (d,3H).

Using the similar procedure the following compounds (Compound ID 2-17)were made.

Example 2(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 2)

LC-MS (ESI): m/z (M+1) 481.1; (M⁺−1) 479.3.

Example 3(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide(Compound ID 3)

LC-MS (ESI): m/z (M+1) 467.2; ¹H NMR (400 MHz, DMSO) δ 10.87 (s, 1H),8.45-8.39 (m, 1H), 8.31 (d, J=4.0 Hz, 1H), 8.22 (t, J=8.2 Hz, 3H),7.92-7.85 (m, 1H), 7.80 (d, J=8.3 Hz, 2H), 7.20 (dd, J=6.9, 5.2 Hz, 1H),5.55 (s, 1H), 4.16-3.51 (m, 4H), 2.46 (dd, J=13.7, 7.1 Hz, 2H),2.06-1.96 (m, 3H).

Example 4(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide(Compound ID 4)

LC-MS (ESI): m/z (M+1) 509.2; (M⁺−1) 507.3

Example 5(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide(Compound ID 5)

LC-MS (ESI): m/z (M+1) 492.1; (M⁺−1) 490.2

Example 6(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide(Compound ID 6)

LC-MS (ESI): m/z (M+1) 523.3; (M⁺−1) 521.3

Example 7(R)-4-(4-amino-1-(1-pent-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 7)

To the solution of(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamidehydrochloride (40 mg, 0.08 mmol) and 2-pentynoic acid (Aldrich, 30 mg,0.30 mmol) in DMF (3 mL) were added diisopropylethylamine (DIEA, 180 μL,1.0 mmol) and then PyBOP (105 mg, 0.20 mmol). The mixture was stirred atRT for 40 min, and treated with TFA (300 μL). It was diluted with 1 mLwater, stirred, and directly subjected to reverse phase preparative HPLCto isolate(R)-4-(4-amino-1-(1-pent-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamideas HCl salt (29 mg).

LC-MS (ESI): m/z (M+H)⁺ 495.2, (M−H)⁻ 493.3. UV: λ=301 nm.

Example 8(R)-4-(4-amino-1-(1-(3-cyclopropylpropioloyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 8)

In a similar manner as described in Example 7,(R)-4-(4-amino-1-(1-(3-cyclopropylpropioloyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamidewas prepared using 3-cyclopropyl-2-propynoic acid (Aldrich). LC-MS(ESI): m/z (M+H)⁺ 507.2, (M−H)⁻ 505.2. UV: λ=301 nm.

Example 9(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide(Compound ID 9)

LC-MS (ESI): m/z (M+1) 509.2; (M⁺−1) 507.3

Example 10(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide(Compound ID 10)

LC-MS (ESI): m/z (M+1) 481.1

Example 11(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 11)

LC-MS (ESI): m/z (M+1) 495.1

Example 12(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide(Compound ID 12)

LC-MS (ESI): m/z (M+1) 549.2

Example 13(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide(Compound ID 13)

LC-MS (ESI): m/z (M+1) 523.2

Example 14(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide(Compound ID 14)

LC-MS (ESI): m/z (M+1) 506.1

Example 15(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide(Compound ID 15)

LC-MS (ESI): m/z (M+1) 473.1; (M⁺−1) 471.2

Example 16(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide(Compound ID 16)

LC-MS (ESI): m/z (M+1) 487.0; (M⁺−1) 485.0.

Example 17(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide(Compound ID 17)

LC-MS (ESI): m/z (M+1) 501.1; (M⁺−1) 499.2

Example 18 Preparation of(R)-4-(4-amino-1-(1-(4-methoxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide(Compound ID 18)

Preparation of 4-methoxybut-2-ynoic acid

To a clear colorless solution of 3-methoxyprop-1-yne (1.0 ml, 11.8 mmol)in 200 ml anhydrous THF, under nitrogen and cooled to −78° C., was addedn-buthyllithium (2.5 M solution in hexanes) (8.26 ml, 20.65 mmol). Theresulting solution was stirred for 20 minutes at −78° C., and then acube of dry ice quickly crushed with mortar and pestle (˜10 g, 236 mmol)was added and allowed to dissolve. The reaction was continued foradditional 30 minutes, then quenched slowly with water and allowed tocome to room temperature. The mixture was diluted with ethyl acetate andwater (pH=6), the pH of which was adjusted to pH=3 using 2M HCl. Theaqueous was extracted twice with ethyl acetate and the organics driedover Na₂SO₄, filtered, concentrated and briefly vacuum-dried.4-Methoxybut-2-ynoic acid (2.08 g) was obtained as a dark yellow liquid,LC-MS (ESI): m/z, (M−H)⁻, 113.0, and used without purification.

Preparation of(R)-4-(4-amino-1-(1-(4-methoxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide(Compound ID 18)

The solution of crude 4-methoxybut-2-ynoic acid (112 mg, ˜0.97 mmol),(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide(100 mg, 0.196 mmol), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide(93.9 mg, 0.490 mmol) and TEA (0.109 ml, 0.784 mmol), in DCM (3.0 ml)was stirred for 16 hours, then extracted with ethyl acetate and dilutedbrine mixture. The aqueous was re-extracted with ethyl acetate and thecombined organics dried over Na₂SO₄, filtered, concentrated and purifiedby reverse-phase HPLC (5 mM HCl aqueous and AcN used as mobile phase).The collected product fractions were frozen and lyophilized to yield(R)-4-(4-amino-1-(1-(4-methoxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamideas a white powder (18.8 mg, 19% yield); LC-MS (ESI): m/z (M+H)⁺, 497.3.

Example 19 Preparation of(R)-4-(4-amino-1-(1-(4-hydroxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide(Compound ID 19)

Preparation of tert-butyldimethyl(prop-2-ynyloxy)silane

To a cloudy mixture of imidazole (6.43 g, 94.5 mmol),tert-butyldimethylchlorosilane (12.96 g, 85.9 mmol) in 140 ml DCM wasadded propargyl alcohol (5 ml, 85.9 mmol) dropwise at room temperature.The resulting mixture was stirred for 60 hours and then filtered thougha pad of silica (10×6 cm), pre-rinsed with heptanes. The residue waswashed with 5% ethyl acetate in heptanes solvent, in portions. Thefiltrate volume was reduced to about 300 ml and then washed with 100 mlwater. The organics were dried over MgSO₄, filtered and concentrated to12.7 g crude (87% yield) tert-butyldimethyl(prop-2-ynyloxy)silane, whichwas used without purification.

Preparation of 4-(tert-butyldimethylsilyloxy)but-2-ynoic acid

Following the procedure described in Example 18 for preparation of4-methoxybut-2-ynoic acid, 4-(tert-butyldimethylsilyloxy)but-2-ynoicacid (1.95 g, 88% yield) was obtained as clear yellow liquid, LC-MS(ESI): m/z, (M−H)⁻, 213.0, and was used without purification.

Preparation of(R)-4-(4-amino-1-(1-(4-hydroxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide

The TBDMS-protected title compound was prepared as Example “1.2Preparation of(R)-4-(4-amino-1-(1-(4-methoxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide”and the crude material purified by Flash Silica Chromatography(Methanol/DCM 0-10% gradient used), upon which the TBDMS-protectinggroup falls off. The title compound (35.7 mg, 38% yield) was re-purifiedusing reverse-phase HPLC and lyophilized to yield 29.2 mg of the whitepowder; LC-MS (ESI): m/z (M+1)⁺, 483.3.

Example 20(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 20)

To the solution of(R)-4-(4-amino-1-(pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(52.4 mg, 0.1 mmol) and DIPEA (77.5 mg, 0.6 mmol) in THF (2 ml) wasadded acryloyl chloride (11.0 mg, 0.12 mmol) at −30° C. After stirred at−30° C. for 30 min, the reaction mixture was stirred at room temperaturefor another 2 hours. Quenched with brine, the aqueous layer wasextracted with EA (3×20 ml). The solvent was evaporated under reducedpressure, the residue was purified by preparative HPLC with 5 mM HCl/ACNas mobile phase to give the desired product:(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(23.8 mg, 47% yield). LC-MS (ESI): m/z (M⁺+1) 469.1; (M⁺−1) 467.2.

The other acryloyl or propenoyl compounds of the invention can be orwere prepared using the method described in Example 20 and using theappropriate α,β-unsaturated acid chloride.

Example 21(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide(Compound ID 21)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1); (M⁺−1) 521.2

Example 22(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide(Compound ID 22)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 455.05; (M⁺−1) 453.2

Example 23(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide(Compound ID 23)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 497.15; (M⁺−1) 495.25

Example 24(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide(Compound ID 24)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 480.05; (M⁺−1) 478.2

Example 25(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide(Compound ID 25)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 469.1; (M⁺−1) 467.2

Example 26(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 26)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 483.1

Example 27(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide(Compound ID 27)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 537.1; (M+−1) 535.3

Example 28(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide(Compound ID 28)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 511.2; (M⁺−1) 509.3

Example 29(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide(Compound ID 29)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 494.1

Example 30(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide(Compound ID 30)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 497.2

Example 31(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide(Compound ID 31)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 511.2; (M⁺−1) 509.3

Example 32(R,E)-4-(4-amino-1-(1-but-2-enoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 32)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+H)⁺ 483.1, (M−H)⁻ 481.3. UV: λ=301 nm.

Example 33(R)-4-(4-amino-1-(1-(3-methylbut-2-enoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 33)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+H)⁺ 497.2, (M−H)⁻ 495.3. UV: λ=301 nm.

Example 34(R,E)-4-(4-amino-1-(1-pent-2-enoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 45)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+H)⁺ 497.1, (M−H)⁻ 495.3. UV: λ=301 nm.

Example 35(R)-4-(4-amino-1-(1-methacryloylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 46)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+H)⁺ 483.1, (M−H)⁻ 481.3. UV: λ=301 nm.

Example 36(R,Z)-4-(4-amino-1-(1-(2-methylbut-2-enoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 47)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+H)⁺ 497.1, (M−H)⁻ 495.3. UV: λ=301 nm.

Example 37(R)-4-(4-amino-1-(1-(2-cyano-3-methylbut-2-enoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 48)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+H)⁺ 522.1, (M−H)⁻ 520.3. UV: λ=301 nm.

Example 38(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide(Compound ID 34)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M+1) 475.1; (M⁺−1) 473.2

Example 39(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide(Compound ID 39)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M⁺−1) 459.2

Example 404-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide(Compound ID 36)

LC-MS (ESI): m/z (M⁺+1) 563.2; (M⁺−1) 561.2

Example 414-(1-((1R,4R)-4-acrylamidocyclohexyl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 37)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M⁺+1) 497.2; (M⁺−1) 495.2

Example 424-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide(Compound ID 38)

LC-MS (ESI): m/z (M⁺+1) 495.2; (M′-1) 493.2

Example 434-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide(Compound ID 39)

LC-MS (ESI): m/z (M⁺+1) 509.2; (M⁺−1) 507.3

Example 444-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide(Compound ID 40)

LC-MS (ESI): m/z (M⁺+1) 537.2; (M⁺−1) 535.3

Example 454-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide(Compound ID 41)

LC-MS (ESI): m/z (M⁺+1) 520.2; (M⁺−1) 518.3

Example 464-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide(Compound ID 42)

LC-MS (ESI): m/z (M⁺+1) 512.2; (M⁺−1) 513.3

Example 474-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide(Compound ID 43)

LC-MS (ESI): m/z (M⁺+1) 501.1; (M⁺−1) 499.2

Example 484-(1-((1R,4R)-4-acrylamidocyclohexyl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide(Compound ID 44)

Prepared following the method described in Example 20.

LC-MS (ESI): m/z (M⁺+1) 489.1; (M⁺−1) 487.2

Example 49(R)-4-(4-amino-1-(1-(4-hydroxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide(Compound ID 49)

LC-MS (ESI): m/z (M⁺+1) 551.4; (M⁺−1) 549.2

Example 50(R)-4-(4-amino-1-(1-(4-methoxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide(Compound ID 50)

LC-MS (ESI): m/z (M⁺+1) 565.4; (M⁺−1) 563.2

Example 51(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(3-methylisothiazol-5-yl)benzamide(Compound ID 51)

51.1 The Preparation of4-(3-methylisothiazol-5-ylcarbamoyl)phenylboronic acid

To a solution of 3-methylisothiazol-5-amine (150 mg, 1 mmol) andpyridine (156 mg, 2 mmol) in dionane was added4-(chlorocarbonyl)phenylboronic acid in small portions. The reactionmixture was stirred overnight at room temperature. The reaction wasquenched with water, extracted with ethyl acetate 3 times. After dryingwith MgSO₄, solvent was removed under reduced pressure and the residuewas used directly for the next step without further purification. LC-MS(ESI): m/z (M+1) 263.2.

51.2 The preparation of(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(3-methylisothiazol-5-yl)benzamide

To a pressure reaction vessel were added(R)-1-(3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidin-1-yl)prop-2-en-1-one(100 mg, 0.25 mmol), 4-(3-methylisothiazol-5-ylcarbamoyl)phenylboronicacid (prepared from 1.1), Pd(dppf)Cl₂ (20 mg, 0.025 mmol), Cs₂CO₃ (162mg, 0.5 mmol), dioxane (4 ml) and water (1 ml). The reaction mixture wasdegassed with N₂ for 2 min. and was heated at 100° C. for 15 min. in asealed tube. After cooling to room temperature, the solvent was removedunder reduced pressure and the residue was purified by prep HPLC to give20.5 mg product. LC-MS (ESI): m/z (M+1) 489.2.

Example 52(R)-4-(4-amino-1-(1-(1-cyanocyclopropanecarbonyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide(Compound ID 52)

UV=295 nm, LC-MS (ESI): m/z (M⁺+1) 494.3; (M⁺−1) 492.1.

Additional Exemplary Compounds of the Invention

Other compounds of the invention have been or can be prepared accordingto the synthetic methods, or some variations thereof, described herein.The compounds can be prepared from readily available starting materialsusing the following general methods and procedures. It will beappreciated that where typical or preferred process conditions (i.e.,reaction temperatures, times, mole ratios of reactants, solvents,pressures, etc.) are given; other process conditions can also be usedunless otherwise stated. Optimum reaction conditions may vary with theparticular reactants or solvent used, but such conditions can bedetermined by one skilled in the art by routine optimization procedures.

Example 53a Btk In Vitro Inhibitory Activity (Method A)

The Btk IC₅₀s of compounds disclosed herein is determined in both acellular kinase assay and in a cellular functional assay of BCR-inducedcalcium flux as described below.

Btk kinase activity is determined using a time-resolved fluorescenceresonance energy transfer (TR-FRET) methodology. Measurements areperformed in a reaction volume of 50 μL using 96-well assay plates.Kinase enzyme, inhibitor, ATP (at the K_(m) for the kinase), and 1 μMpeptide substrate (Biotin-AVLESEEELYSSARQ-NH₂) are incubated in areaction buffer composed of 20 mM Tris, 50 mM NaCl, MgCl₂ (5-25 mMdepending on the kinase), MnCl₂ (0-10 mM), 1 mM DTT, 0.1 mM EDTA, 0.01%bovine serum albumin, 0.005% Tween-20, and 10% DMSO at pH 7.4 for onehour. The reaction is quenched by the addition of 1.2 equivalents ofEDTA (relative to divalent cation) in 25 μL of 1× Lance buffer(Perkin-Elmer). Streptavidin-APC (Perkin-Elmer) and Eu-labeled p-Tyr100antibody (Perkin-Elmer) in 1× Lance buffer are added in a 25 μL volumeto give final concentrations of 100 nM and 2.5 nM, respectively, and themixture is allowed to incubate for one hour. The TR-FRET signal ismeasured on a multimode plate reader with an excitation wavelength(λ_(Ex)) of 330 nm and detection wavelengths (λ_(Em)) of 615 and 665 nm.Activity is determined by the ratio of the fluorescence at 665 nm tothat at 615 nm. For each compound, enzyme activity is measured atvarious concentrations of compound. Negative control reactions areperformed in the absence of inhibitor in replicates of six, and twono-enzyme controls are used to determine baseline fluorescence levels.Inhibition constants, K_(i)(app), ware obtained using the programBatchK_(i) (Kuzmic et al. (2000), Anal. Biochem. 286:45-50). IC₅₀s areobtained according to the equation:

IC ₅₀ ={Ki(app)/(1+[ATP]/K _(m) ^(ATP))}+[E] _(total)/2;

For all kinases, [ATP]=K_(m) ^(ATP), [Btk]_(total)=0.5 nM and[Lck]_(total)=6 nM.

Example 53b Btk In Vitro Inhibitory Activity (Method B)

Kinase activity is measured in vitro using electrophoretic mobilityshift assay. The kinase reactions are assembled in a total volume of 25μL in 384 well plates. The reactions comprise: BTK enzyme (1 nM,N-terminal His6-tagged, recombinant, full-length, human BTK purifiedfrom baculovirus Sf21 insect cell system), inhibitor, ATP (16 μM, theapparent K_(m) for the kinase), fluorescently labeled peptide substrate(1 μM, FAM-GEEPLYWSFPAKKK-NH2) in a reaction buffer composed of 100 mMHEPES, pH7.5, 5 mM MgCl₂ 1 mM DTT, 0.1% bovine serum albumin, 0.01%Triton X-100, and 1% DMSO. The reaction is incubated for one hour and isquenched by the addition of 45 μL of termination buffer (100 mM HEPES,pH7.5, 0.01% Triton X-100, 30 mM EDTA). The terminated reactions areanalyzed using 12 channel LabChip® 3000 microfluidic detectioninstrument (Caliper Life Sciences). The enzymatic phosphorylation of thepeptide results in a change in net charge, enabling electrophoreticseparation of product from substrate peptide. As substrate and productpeptides are separated, two peaks of fluorescence are observed. Changein the relative fluorescence intensity of the substrate and productpeaks is the parameter measured, reflecting enzyme activity. In thepresence of an inhibitor, the ratio between product and substrate isaltered:the signal of the product decreases, while the signal of thesubstrate increases.

Activity in each sample is determined as the product to sum ratio(PSR):P/(S+P), where P is the peak height of the product peptide and Sis the peak height of the substrate peptide. For each compound, enzymeactivity is measured at various concentrations (12 concentrations ofcompound spaced by 3× dilution intervals). Negative control samples(0%−inhibition in the absence of inhibitor) and positive control samples(100%−inhibition, in the presence of 20 mM EDTA) are assembled inreplicates of four and are used to calculate %−inhibition values foreach inhibitor at each concentration. Percent inhibition (P_(inh)) isdetermined using following equation:

P_(inh)=(PSR_(0%)−PSR_(inh))/(PSR_(0%)−PSR_(100%))*100, where PSR_(inh)is the product sum ratio in the presence of inhibitor, PSR_(0%) is theaverage product sum ration in the absence of inhibitor and PSR_(100%) isthe average product sum ratio in 100%−inhibition control samples;

The IC₅₀ values of inhibitors are determined by 4 parameter sigmoidaldose-response model fitting of the inhibition curves (P_(inh) versusinhibitor concentration) using XLfit 4 software.

Example 53c Btk In Vitro Inhibitory Activity (Method C)

Human Btk kinase (Genbank accession # NP_(—)000052) was purified frominsect cells as a full-length construct containing a N-terminal 6×-Histag. Btk kinase activity was determined using a radiometric filterbinding assay. Measurements are performed in a low μL reaction volume384-well assay plates. BTK enzyme (8 nM final in reaction), inhibitor(at requested doses), and 0.2 mg/mL peptide substrate (Poly-Glu-Tyr, 4:1ratio) are incubated in a reaction buffer composed of 20 mM Hepes (pH7.5), 10 mM MgCl₂, 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mMNa₃VO₄, 2 mM DTT, 1% DMSO for 15 min. followed by addition of 1 μM ATPto start the assay. Kinase reactions are carried out for 120 min. atroom temperature. The reaction was stopped by spotting of reactionsample onto P81 cationic exchange paper (Whatman). Unbound phosphate wasremoved by extensive washing of filters in 0.75% Phosphoric acid. Aftersubtraction of background derived from control reactions containinginactive enzyme (via addition of saturating EDTA), kinase activity datafor each dose of compound tested was expressed as the percent ofremaining kinase activity in test samples compared to vehicle (dimethylsulfoxide) reactions. IC₅₀ values and curve fits were obtained usingPrism (GraphPad Software).

The degree of Btk inhibition was determined using one of the methodsoutlined in Example 53a, 53b and 53c.

TABLE 1 IC₅₀ Values for Exemplary Compounds of the Invention Compound IDName Btk IC₅₀ 1(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d] Apyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide 2(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 3(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide 4(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide 5(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide 6(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide 7(R)-4-(4-amino-1-(1-pent-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 8(R)-4-(4-amino-1-(1-(3-cyclopropylpropioloyl)pyrrolidin-3-yl)-1H- Apyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 9(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide 10(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide 11(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 12(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide 13(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide 14(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide 15(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide 16(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide 17(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide 20(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 21(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide 22(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide 23(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide 24R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide 25(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide 26(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 27(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide 28(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide 29(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide 30(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide 31(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide 32(R,E)-4-(4-amino-1-(1-but-2-enoylpyrrolidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 33(R)-4-(4-amino-1-(1-(3-methylbut-2-enoyl)pyrrolidin-3-yl)-1H- Bpyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 34(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide 35(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide 364-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide 374-(1-((1R,4R)-4-acrylamidocyclohexyl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 384-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide 394-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 404-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide 414-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide 424-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide 434-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide 444-(1-((1R,4R)-4-acrylamidocyclohexyl)-4-amino-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide 45(R,E)-4-(4-amino-1-(1-pent-2-enoylpyrrolidin-3-yl)-1H-pyrazolo[3,4- Ad]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 46(R)-4-(4-amino-1-(1-methacryloylpyrrolidin-3-yl)-1H-pyrazolo[3,4- Bd]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 47(R,Z)-4-(4-amino-1-(1-(2-methylbut-2-enoyl)pyrrolidin-3-yl)-1H- Bpyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 48(R)-4-(4-amino-1-(1-(2-cyano-3-methylbut-2-enoyl)pyrrolidin-3-yl)- B1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide 49(R)-4-(4-amino-1-(1-(4-hydroxybut-2-ynoyl)pyrrolidin-3-yl)-1H- Apyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide 50(R)-4-(4-amino-1-(1-(4-methoxybut-2-ynoyl)pyrrolidin-3-yl)-1H- Apyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide 52(R)-4-(4-amino-1-(1-(1-cyanocyclopropanecarbonyl)pyrrolidin-3-yl)- B1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide IC₅₀: A ≦ 10nM; B > 10 nM

Example 54 Inhibition of a Panel of Kinases

The degree of inhibition of a panel of kinases is determined usingassays similar to the kinase assay described in Example 53.

TABLE 2 IC₅₀ Values for Exemplary Compounds of the Invention Compd IDBtk (IC₅₀) EGFR (IC₅₀) LCK (IC₅₀) JAK3 (IC₅₀) 1 A C C D 2 A D C D 3 A CB D 4 A D C D 5 A D C D 6 A D C D 7 A D D D 8 A C D D 9 A A C — 10 A A C— 11 A B B — 12 A C B — 13 A C A — 14 A C B — 15 A A C — 16 A B B — 17 AA C — 18 A A B — 19 A B C — 20 A A B — 21 A A B — 22 A A C — 52 C — — —IC₅₀: A ≦ 10 nM; 10 nM < B ≦ 100 nM; 100 nM < C ≦ 1 μM; D > 1 μM; — = nodata

Example 55 Lymphoma Cell Assay

Cells of lymphoma cell line DOHH2 were seeded in 96-well plates instandard growth media (RPMI+10% fetal calf serum). Compounds were addedin an 8-point dilution series ranging from 10 to 0.004 μM with DMSO at0.1% final concentration in all wells. Cells were then stimulated with aF(ab′)₂ fragment of anti-human IgG at 10 μg/mL for 14 to 20 hours beforestaining for FACS analysis. Cells were incubated withfluorescent-labeled anti-CD69 antibody for 30 minutes on ice in FACSbuffer and analyzed on the FACSCanto flowcytometry. The IC₅₀, theconcentration that results in a 50% decrease in intensity offluorescence detected on the cell surface, was calculated using thePrizm software to fit the dose-response curve.

Exemplifying compounds, including Compound IDs 20, 21, 23, 26, 27, 30,and 31, were tested in this assay and displayed IC₅₀ values of less than5 nM.

Example 56 Splenocyte Btk Binding Analysis

Spleens were harvested from rats four hours post dose with a compound ofthe invention at 3 mg/kg or 12 mg/kg or vehicle. Splenocytes weresuspended in DPBS. Covalent active site binding to splenic Btk wasdetermined ex vivo by measuring the exclusion of a fluorescent probe(U.S. Pat. No. 8,883,435). Splenic Btk was labeled with the fluorescentprobe and Btk labeling was quantified by SDS-PAGE gel analysis followedby scanning with a Typhoon gel imager. The splenocyte cell suspensionwas lysed by freeze-thawing the cell suspension in liquid nitrogen for 2minutes and thawing in a 37° C. water bath for 5 minutes. Thefreeze/thaw cycle was repeated a total of 4 times. At the completion ofthe freeze/thaw cycles, the splenocytes were spun at maximum speed(13,200 rpm) for 10 minutes. After centrifugation, the supernatant wascollected into a clean 1.5 ml microfuge tube. 1× of 100× stock proteaseinhibitor (Sigma P8340) was added to the cell lysates. The cell lysateswere quantified using a BCA Protein Assay (Pierce). Lysates were dilutedin D-PBS to a final concentration of 1.6 mg/ml followed by incubationwith 2.5 μM of the Btk probe in a 37° C. water bath for 1 hour. Theprobe labeling was halted by the addition of a LDS sample buffer andsample reducing agent (Invitrogen). The samples were then heated to 70°C. for 10 minutes prior to gel electrophoresis. Each spleen was analyzedby adding 25 μl of each sample on a NuPAGE Novex 4-12% Bis-Tris gel(Invitrogen) in the dark. Bodipy-FL labeled Btk was visualized byscanning the gel with a Molecular Dynamics Typhoon scanner. Gels werethen analyzed by standard western blotting techniques to detect Btkprotein. Btk was confirmed present in all lanes. Band intensity in theprobe gels and western blots were measured using ImageQuant (GE).

Example 57 Pharmaceutical Compositions

The compositions described below are presented with a compound ofFormula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) for illustrative purposes.

Example 57a Parenteral Composition

To prepare a parenteral pharmaceutical composition suitable foradministration by injection, 100 mg of a water-soluble salt of acompound of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) is dissolved inDMSO and then mixed with 10 mL of 0.9% sterile saline. The mixture isincorporated into a dosage unit form suitable for administration byinjection.

Example 57b Oral Composition

To prepare a pharmaceutical composition for oral delivery, 100 mg of acompound of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) is mixed with 750mg of starch. The mixture is incorporated into an oral dosage unit for,such as a hard gelatin capsule, which is suitable for oraladministration.

Example 57c Sublingual (Hard Lozenge) Composition

To prepare a pharmaceutical composition for buccal delivery, such as ahard lozenge, mix 100 mg of a compound of Formula (I), (Ia)-(Ic),(IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh),(VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or(XIIa)-(XIIh) with 420 mg of powdered sugar mixed, with 1.6 mL of lightcorn syrup, 2.4 mL distilled water, and 0.42 mL mint extract. Themixture is gently blended and poured into a mold to form a lozengesuitable for buccal administration.

Example 57d Inhalation Composition

To prepare a pharmaceutical composition for inhalation delivery, 20 mgof a compound of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) is mixed with 50 mgof anhydrous citric acid and 100 mL of 0.9% sodium chloride solution.The mixture is incorporated into an inhalation delivery unit, such as anebulizer, which is suitable for inhalation administration.

Example 57e Rectal Gel Composition

To prepare a pharmaceutical composition for rectal delivery, 100 mg of acompound of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) is mixed with 2.5 gof methylcelluose (1500 mPa), 100 mg of methylparapen, 5 g of glycerinand 100 mL of purified water. The resulting gel mixture is thenincorporated into rectal delivery units, such as syringes, which aresuitable for rectal administration.

Example 57f Topical Gel Composition

To prepare a pharmaceutical topical gel composition, 100 mg of acompound of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) is mixed with 1.75g of hydroxypropyl celluose, 10 mL of propylene glycol, 10 mL ofisopropyl myristate and 100 mL of purified alcohol USP. The resultinggel mixture is then incorporated into containers, such as tubes, whichare suitable for topical administration.

Example 57 g Ophthalmic Solution Composition

To prepare a pharmaceutical ophthalmic solution composition, 100 mg of acompound of Formula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh),(IVa)-(IVd), (Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh),(IXa)-(IXd), (Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) is mixed with 0.9 gof NaCl in 100 mL of purified water and filtered using a 0.2 micronfilter. The resulting isotonic solution is then incorporated intoophthalmic delivery units, such as eye drop containers, which aresuitable for ophthalmic administration.

Example 58 Clinical Trial of the Safety and Efficacy of a Compound ofFormula (I), (Ia)-(Ic), (IIa)-(IId), (IIIa)-(IIIh), (IVa)-(IVd),(Va)-(Vd), (VIa)-(VIh), (VIIa)-(VIId), (VIIIa)-(VIIIh), (IXa)-(IXd),(Xa)-(Xh), (XIa)-(XIj) or (XIIa)-(XIIh) in Rheumatoid Arthritis Patients

The purpose of this study is to determine the safety and efficacy of acompound of Formula (I)-(XId) in patients with rheumatoid arthritis.

Inclusion Criteria

-   -   Adult males/Females aged 18˜80 years.    -   Patients who are taking NSAIDs for the treatment of rheumatoid        arthritis.    -   Patients who belong to ACR functional class 1, 2, 3.

Exclusion Criteria

-   -   Patients who belong to ACR functional class 4.    -   Patients who are hypersensitive to clinical trial medicines or        excipient.    -   Patients who have experience of Cerebrovascular bleeding,        bleeding disorder.

Study Design

-   -   Allocation: Randomized, placebo-controlled.    -   Intervention Model: Single Group Assignment.    -   Masking: Double Blind (Subject, Caregiver).    -   Primary Purpose: Supportive Care

Primary Outcome Measures

-   -   Changes in ‘100 mm pain VAS’ value from baseline [Time Frame:        −14, 0, 14, 28, 42 day][Designated as safety issue: No].

Determine PK of an Orally Administered Compound of Formula (I)-(XIIh).

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference in theirentirety for all purposes.

1. A kinase inhibitor which is a compound according to formula (I):

wherein: A is

Hy is 2-pyridyl substituted with 1-5 groups independently selected fromR⁴, or Hy is imidazolyl, thiazolyl, oxazolyl, pyrazolyl, oxadiazolyl,thiadiazolyl, triazolyl, isothiazolyl, or tetrazolyl, each of whichsubstituted with 1-2 groups independently selected from R⁴; each R¹ andR² is independently H, alkyl, or CN; or R¹ and R² together form a bond;R³ is independently H, alkyl, CN, or cycloalkyl; each R⁴ isindependently H, halo, hydroxyl, CN, substituted or unsubstituted alkyl,substituted or unsubstituted amino, substituted or unsubstituted alkoxy,substituted or unsubstituted amido, substituted or unsubstitutedsulfonyl, substituted or unsubstituted carboxy, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl; or twoadjacent R⁴s connect together with Hy to form a bicyclic ring; each n isindependently 0, 1, or 2; and p is 0, 1 or 2; or a metabolite, asolvate, a pharmaceutically acceptable salt, or a prodrug thereof.
 2. Akinase inhibitor which is a compound according to formula (Ia), (Ib) or(Ic):

wherein Hy, R¹, R², R³, and n are as in claim 1; or a pharmaceuticallyacceptable salt, solvate, or prodrug thereof.
 3. The kinase inhibitoraccording to claim 1, wherein Hy is 2-pyridyl substituted with 1-5groups independently selected from R⁴.
 4. The kinase inhibitor accordingto claim 1, wherein Hy is 2-thiazolyl substituted with 1-2 groupsindependently selected from R⁴.
 5. The kinase inhibitor according toclaim 1, wherein the compound is according to formula (IIa), (IIb),(IIc), or (IId):

wherein R¹, R², R³, R⁴, and n are as in claim 1; and m is 1, 2, or 3; ora pharmaceutically acceptable salt, solvate, or prodrug thereof.
 6. Thekinase inhibitor according to claim 1, wherein R¹ is H, Me, Et, n-Pr,i-Pr, or CN.
 7. The kinase inhibitor according to claim 1, wherein R² isH, Me, Et, n-Pr, or i-Pr.
 8. (canceled)
 9. (canceled)
 10. The kinaseinhibitor according to claim 1, wherein the compound is according toformula (IIIa), (IIIb), (IIIc), or (IIId):

wherein R³, R⁴, and n are as in claim 1; or a pharmaceuticallyacceptable salt, solvate, or prodrug thereof.
 11. The kinase inhibitoraccording to claim 1, wherein the compound is according to formula(IIIe), (IIIf), (IIIg) or (IIIh):

wherein R³, R⁴, and n are as in claim 1; or a pharmaceuticallyacceptable salt, solvate, or prodrug thereof.
 12. (canceled) 13.(canceled)
 14. The kinase inhibitor according to claim 1, wherein thecompound is according to formula (IVa), (IVb), (IVc) or (IVd):

wherein R³, and R⁴ are as in claim 1; or a pharmaceutically acceptablesalt, solvate, or prodrug thereof.
 15. The kinase inhibitor according toclaim 1, wherein the compound is according to formula (Va), (Vb), (Vc)or (Vd):

wherein R³, and R⁴ are as in claim 1; or a pharmaceutically acceptablesalt, solvate, or prodrug thereof.
 16. (canceled)
 17. The kinaseinhibitor according to claim 1, wherein R³ is C₁-C₄ alkyl.
 18. Thekinase inhibitor according to claim 1, wherein R³ is H, Me, Et, n-Pr, ori-Pr.
 19. (canceled)
 20. (canceled)
 21. (canceled)
 22. (canceled) 23.(canceled)
 24. The kinase inhibitor according to claim 1, wherein R³ iscycloalkyl.
 25. The kinase inhibitor according to claim 1, wherein R³ iscyclopropyl.
 26. (canceled)
 27. The kinase inhibitor according to claim1, wherein the compound is according to formula (VIa), (VIb), (VIc), or(VId):

wherein R⁴ is as in claim 1; or a pharmaceutically acceptable salt,solvate, or prodrug thereof.
 28. The kinase inhibitor according to claim1, wherein the compound is according to formula (VIe), (VIf), (VIg) or(VIh):

wherein R⁴ is as in claim 1; or a pharmaceutically acceptable salt,solvate, or prodrug thereof.
 29. The kinase inhibitor according to claim1, wherein the compound is according to formula (VIIa), (VIIb), (VIIc),or (VIId):

wherein R⁴ is as in claim 1; or a pharmaceutically acceptable salt,solvate, or prodrug thereof.
 30. The kinase inhibitor according to claim1, wherein R⁴ is H, Me, Et, n-Pr, i-Pr, CF₃ or CN.
 31. (canceled) 32.(canceled)
 33. (canceled)
 34. (canceled)
 35. The kinase inhibitoraccording to claim 1, wherein the compound is according to formula(VIIIa), (VIIIb), (VIIIc) or (VIIId):

or a pharmaceutically acceptable salt, solvate, or prodrug thereof. 36.The kinase inhibitor according to claim 1, wherein the compound isaccording to formula (VIIIe), (VIIIf), (VIIIg) or (VIIIh):

or a pharmaceutically acceptable salt, solvate, or prodrug thereof. 37.The kinase inhibitor according to claim 1, wherein the compound isaccording to formula (IXa), (IXb), (IXc) or (IXd):

or a pharmaceutically acceptable salt, solvate, or prodrug thereof. 38.The kinase inhibitor according to claim 1, wherein the compound isaccording to formula (Xa), (Xb), (Xc) or (Xd):

or a pharmaceutically acceptable salt, solvate, or prodrug thereof. 39.The kinase inhibitor according to claim 1, wherein the compound isaccording to formula (Xe), (Xf), (Xg), or (Xh):

or a pharmaceutically acceptable salt, solvate, or prodrug thereof. 40.The kinase inhibitor according to claim 1, wherein the compound isaccording to formula (XIa), (XIb), (XIc), or (XId):

or a pharmaceutically acceptable salt, solvate, or prodrug thereof. 41.A kinase inhibitor which is a compound selected from:(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-pent-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-(3-cyclopropylpropioloyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide;(R)-4-(4-amino-1-(1-but-2-ynoylpiperidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide;(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide;R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpiperidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-propylpyridin-2-yl)benzamide;(R,E)-4-(4-amino-1-(1-but-2-enoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-(3-methylbut-2-enoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide;(R)-4-(1-(1-acryloylpyrrolidin-3-yl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide;4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;4-(1-((1R,4R)-4-acrylamidocyclohexyl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-isopropylpyridin-2-yl)benzamide;4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-cyanopyridin-2-yl)benzamide;4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(5-methylthiazol-2-yl)benzamide;4-(4-amino-1-((1R,4R)-4-but-2-ynamidocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide;4-(1-((1R,4R)-4-acrylamidocyclohexyl)-4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(thiazol-2-yl)benzamide;(R,E)-4-(4-amino-1-(1-pent-2-enoylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-methacryloylpyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R,Z)-4-(4-amino-1-(1-(2-methylbut-2-enoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-(2-cyano-3-methylbut-2-enoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-methylpyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-(4-hydroxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-(4-methoxybut-2-ynoyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)benzamide;(R)-4-(4-amino-1-(1-(1-cyanocyclopropanecarbonyl)pyrrolidin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-N-(pyridin-2-yl)benzamide;and a solvate or a pharmaceutically acceptable salt thereof. 42.(canceled)
 43. A pharmaceutical composition comprising a kinaseinhibitor of claim 1; or a pharmaceutically acceptable salt, solvate, orprodrug thereof; and a pharmaceutically acceptable excipient. 44.(canceled)
 45. A method for treating a disease or condition that iscausally related to the aberrant activity of a tyrosine kinase receptorcomprising administering to a patient in need thereof; a therapeuticallyeffective amount of the pharmaceutical composition of claim
 43. 46.(canceled)
 47. (canceled)
 48. (canceled)
 49. (canceled)
 50. (canceled)51. (canceled)
 52. (canceled)
 53. (canceled)
 54. (canceled) 55.(canceled)
 56. (canceled)
 57. (canceled)
 58. (canceled)
 59. (canceled)60. (canceled)
 61. (canceled)
 62. (canceled)
 63. (canceled) 64.(canceled)
 65. (canceled)
 66. (canceled)